JP2020011984A - Combination of anti-lag-3 antibodies and anti-pd-1 antibodies for treating tumors - Google Patents

Abstract

To provide methods for treatment of solid tumors with very poor prognosis (e.g., advanced refractory solid tumors).SOLUTION: The method comprises intravenously administering an anti-lymphocyte activation gene-3 antibody in combination with an anti-programmed cell death-1 antibody, according to a particular clinical dosage regimen, i.e., at particular doses and according to a specific dosing schedule, thereby producing at least one therapeutic effect selected from among a reduction in size of a tumor, a reduction in the number of metastatic lesions over time, a complete response, a partial response, and a stable disease.SELECTED DRAWING: None

Description

RELATED APPLICATIONS This application is related to U.S. Provisional Patent Application No. 61 / 880,606 filed September 20, 2013 and U.S. Provisional Patent Application No. 62 / 014,471 filed June 19, 2014. Claim priority. The contents of any patents, patent applications, and references cited throughout this specification are hereby incorporated by reference in their entirety.

Lymphocyte activating gene-3 (LAG-3; CD223) is a type I transmembrane protein expressed on the cell surface of activated CD4 ⁺ and CD8 ⁺ T cells and a subset of NK cells and dendritic cells (Triebel F, et al., J. Exp. Med. 1990; 171: 1393-1405; Workman CJ, et al., J. Immunol. 2009; 182 (4): 1885-91). LAG-3 is closely related to CD4, a co-receptor for helper T cell activation. Both molecules have four extracellular Ig-like domains and require binding to their ligand, the major histocompatibility complex (MHC) class II, for their functional activity. In contrast to CD4, LAG-3 is expressed only on the cell surface of activated T cells, and its cleavage from the cell surface terminates LAG-3 signaling. LAG-3 is also found as a soluble protein, but does not bind to MHC class II and its function is unknown.

LAG-3 has been reported to play an important role in promoting regulatory T cell (Treg) activity and in negatively regulating T cell activation and proliferation (Workman CJ, et al., J. Immunol. 2005). 174: 688-695). Both native and derived Tregs have elevated LAG-3 expression, which is required for Tregs to exert their suppressive function to the full (Camisaschi C, et al., J. Immunol. 2010; 184: 6545-6551 and Huang CT, et al., Immunity. 2004; 21: 503-513). Furthermore, ectopic expression of LAG-3 on CD4 ⁺ effector T cells reduced its proliferative capacity and conferred them the ability to regulate third party T cells (Huang CT, et al., Immunity. 2004; 21: 503-513). Also, recent studies have shown that high LAG-3 expression on depleted lymphocytic choriomeningitis virus (LCMV) -specific CD8 ⁺ T cells contributes to its unresponsive state, and CD8 ⁺ T cell antitumor response (Blackburn SD, et al., Nat. Immunol. 2009; 10: 29-37 and Grosso JF, et al., J. Clin. Invest. 2007; 117: 3383-3392). In fact, LAG-3 maintained tolerance to self and tumor antigens in two mouse models by a direct effect on CD8 ⁺ T cells (Grosso JF, et al., J. Clin. Invest. 2007; 117: 3383). -3392).

  However, the immune tolerance observed in the context of tumor development and tumor recurrence is thought to be mediated not only by LAG-3 but also by co-expression of various T cell negative regulatory receptors. Chronic virus infection model (Blackburn SD, et al., Nat. Immunol. 2009; 10: 29-37, Grosso JF, et al., J. Clin. Invest. 2007; 117: 3383-3392, and Lyford-Pike S , et al., Cancer Res. 2013; 73 (6): 1733-41), knockout mice (Woo SR, et al., Cancer Res. 2012; 72: 917-927; Okazaki T, et al., J. Exp Med. 2011; 208: 395-407, and Bettini M. et al., J. Immunol. 2011; 187: 3493-3498), tumor recurrence model (Goding SR, et al., J. Immunol. 2013; 190) (9): 4899-4909) and, to a more limited extent, human cancer patients (Goding SR, et al., J. Immunol. 2013; 190 (9): 4899-4909, Matsuzaki J, et al. Natl. Acad. Sci., US A. 2010; 107: 7875-7880, and Gandhi MK, et al., Blood. 2006; 108: 2280-2289). It supports a model in which exposed T cells are progressively inactivated by a process called "depletion." Exhausted T cells are characterized by expression of T cell negative regulatory receptors, mainly CTLA-4, PD-1, and LAG-3, whose actions are to proliferate, produce cytokines, and target cells. To limit the ability of cells to kill and / or increase Treg activity. However, the timing and order of expression of these molecules in tumor development and recurrence has not been fully characterized.

  Programmed Cell Death 1 (PD-1: Programmed Cell Death 1) is a cell surface signaling receptor that plays a critical role in regulating T cell activation and tolerance (Keir ME, et al., Annu Rev Immunol) 2008; 26: 677-704). It is a type I transmembrane protein and, together with BTLA, CTLA-4, ICOS and CD28, constitutes the T cell costimulatory receptor of the CD28 family. PD-1 is mainly expressed on activated T cells, B cells, and myeloid cells (Dong H, et al., Nat Med. 1999; 5: 1365-1369). It is also expressed on natural killer (NK) cells (Terme M, et al., Cancer Res 2011; 71: 5393-5399). Binding of PD-1 with its ligands PD-L1 and PD-L2 phosphorylates tyrosine residues in the adjacent intracellular immune receptor tyrosine inhibition domain, and then recruits phosphatase SHP-2, Eventually, T cell activation is down-regulated. One important role of PD-1 is to limit the activation of T cells in peripheral tissues at the time of the inflammatory response to infection, thus limiting the development of autoimmunity (Pardoll DM., Nat Rev Cancer). 2012; 12: 252-264). Evidence for this negative regulatory role comes from the finding that PD-1 deficient mice develop cardiomyopathy along with lupus-like autoimmune diseases, including arthritis and nephritis (Nishimura H, et al., Immunity, 1999). 11: 141-151 and Nishimura H, et al., Science, 2001; 291: 319-322). In the context of a tumor, the result is the development of immune tolerance within the tumor microenvironment. PD-1 is highly expressed on tumor-infiltrating lymphocytes and its ligand is up-regulated on the cell surface of many different tumors (Dong H, et al., Nat Med 2002; 8: 793-800). Multiple mouse cancer models have shown that binding of ligand to PD-1 results in immune evasion. Furthermore, blocking this interaction results in antitumor activity (Topalian SL, et al. NEJM 2012; 366 (26): 2443-2454; Hamid O, et al., NEJM 2013; 369: 134-144). Furthermore, inhibition of the PD-1 / PD-L1 interaction has been shown to mediate potent antitumor activity in preclinical models (U.S. Pat. Nos. 8,008,449 and 7,943,743). ).

  Patients with metastatic or refractory solid tumors have very poor prognosis [Rosenberg SA, et al., Cancer immunotherapy in Cancer: Principles & Practice of Oncology (Eds DeVita VT, Lawrence TS and Rosenberg SA) 2011; 332- 344 (Lippincott Williams & Wilkins, Philadelphia PA)]. Despite advances in various therapies, the increase in overall survival in this patient population was limited. Accordingly, it is an object of the present invention to provide an improved method for treating a subject having such a tumor (eg, an advanced refractory solid tumor).

  A method for treating a tumor in a human patient, particularly a solid tumor (eg, an advanced refractory solid tumor), comprising administering to the patient a combination of an anti-LAG-3 antibody and an anti-PD-1 antibody. Methods are provided herein, including, and wherein the combination is administered (or is for administration) according to a particular clinical dosing regimen (ie, at a particular dose, according to a particular dosing schedule). In one embodiment, the human patient has melanoma, non-small cell lung cancer (NSCLC), virus-related cancer, head and neck cancer (NHC), or gastric adenocarcinoma.

  An exemplary anti-LAG-3 antibody is BMS-986016 comprising a heavy and light chain comprising the sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively, or an antigen-binding fragment and variant thereof. In another embodiment, the antibody comprises the heavy and light chain complementarity determining regions (CDRs) or variable regions (VRs) of BMS-986016. Thus, in one embodiment, the antibody comprises a CDR1, CDR2, and CDR3 domain of the heavy chain variable (VH) region of BMS-986016 having the sequence set forth in SEQ ID NO: 3, and a BMS having the sequence set forth in SEQ ID NO: 5. Includes the CDR1, CDR2 and CDR3 domains of the light chain variable (VL) region of -9886016. In another embodiment, the antibody is a CDR1, CDR2, and CDR3 heavy chain sequence set forth in SEQ ID NOs: 7, 8, and 9, respectively, and a CDR1, CDR2, and CDR3 light chain sequence set forth in SEQ ID NOs: 10, 11, and 12, respectively. including. In another embodiment, the antibody has a VH and / or VL region comprising the amino acid sequence set forth in SEQ ID NO: 3 and / or SEQ ID NO: 5, respectively. In another embodiment, the antibody comprises the VH and / or VL regions encoded by the nucleic acid sequences set forth in SEQ ID NO: 4 and / or SEQ ID NO: 6, respectively. In another embodiment, the antibody competes for and / or binds to the same epitope on LAG-3 as the antibody described above. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with an antibody described above (eg, at least about 90%, 95% or 99% variable amino acid identity with SEQ ID NO: 3 or SEQ ID NO: 5). Region identity).

  An exemplary anti-PD-1 antibody is nivolumab (also referred to as “5C4” in WO2006 / 121168, including BMS-936558, MDX) comprising a heavy and light chain comprising the sequences set forth in SEQ ID NO: 17 and SEQ ID NO: 18, respectively. -1106 and ONO-4538), or antigen-binding fragments and variants thereof. In other embodiments, the antibody comprises the heavy and light chain CDRs or VRs of BMS-936558. Thus, in one embodiment, the antibody comprises a CDR1, CDR2 and CDR3 domain of the VH region of BMS-936558 having the sequence set forth in SEQ ID NO: 19, and a VL region of BMS-936558 having the sequence set forth in SEQ ID NO: 21. CDR1, CDR2 and CDR3 domains. In another embodiment, the antibody is a heavy chain CDR1, CDR2 and CDR3 domain comprising the sequence set forth in SEQ ID NOs: 23, 24 and 25, respectively, and a light chain comprising the sequence set forth in SEQ ID NOs: 26, 27 and 28, respectively. Includes CDR1, CDR2 and CDR3 domains. In another embodiment, the antibody comprises a VH and / or VL region comprising the amino acid sequence set forth in SEQ ID NO: 19 and / or SEQ ID NO: 21, respectively. In another embodiment, the antibody comprises a variable heavy (VH) and / or variable light (VL) region encoded by the nucleic acid sequence set forth in SEQ ID NO: 20 and / or SEQ ID NO: 22, respectively. In another embodiment, the antibody competes for and / or binds to the same epitope on PD-1 as the antibody described above. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with the antibody described above (eg, at least about 90%, 95%, or 99% of SEQ ID NO: 19 or SEQ ID NO: 21). Variable region identity).

Accordingly, in one aspect, a method of treating a solid tumor (eg, an advanced refractory solid tumor) in a human patient, comprising:
(A) an anti-LAG- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 5; 3 antibodies,
(B) an anti-PD- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 19, and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 21; Administering an effective amount of each of the one antibody,
Including at least one dosing cycle, the cycle is eight weeks, and for at least one of each cycle, four doses of anti-LAG-3 antibody are administered at a dose of 3, 20, 80, or 240 mg, and four doses of anti-PD- A method is provided wherein one antibody is administered at a dose of 80 or 240 mg. In another embodiment, four doses of the anti-LAG-3 antibody are administered at a dose of about 0.03, 0.25, 1 or 3 mg / kg body weight, and four doses of the anti-PD-1 antibody are 1 or It is administered at a dose of 3 mg / kg body weight.

In one embodiment, the anti-LAG-3 and anti-PD-1 antibodies are:
(A) 3 mg of anti-LAG-3 antibody and 80 mg of anti-PD-1 antibody;
(B) 3 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(C) 20 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(D) 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(E) Administered at a dose of 240 mg anti-LAG-3 antibody and 240 mg anti-PD-1 antibody.

In another embodiment, the anti-LAG-3 and PD-1 antibodies are
(A) 0.03 mg / kg anti-LAG-3 antibody and 1 mg / kg anti-PD-1 antibody;
(B) 0.03 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody;
(C) 0.25 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody;
(D) 1 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody; or (e) administered at a dose of 3 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody Is done.

  In one embodiment, the dose of the anti-LAG-3 and / or anti-PD-1 antibodies is calculated per mg / kg body weight. In another embodiment, the dose of the anti-LAG-3 antibody and / or anti-PD-1 antibody is a fixed fixed dose. In another embodiment, an intermediate dose of LAG-3 and / or PD-1 is used. For example, LAG-3 can be administered at 0.4 mg / kg and PD-1 can be administered at 90 mg / kg. In another embodiment, the dosing regimen is adjusted to provide the optimal desired response (eg, an effective response).

  In another embodiment, the anti-PD-1 antibody is administered on days 1, 15, 29 and 43 of each cycle. In another embodiment, the anti-LAG-3 antibody is administered on days 1, 15, 29 and 43 of each cycle. In another embodiment, the anti-PD-1 antibody is administered prior to the administration of the anti-LAG-3 antibody. In another embodiment, the anti-PD-1 antibody is administered after administration of the anti-LAG-3 antibody. In another embodiment, the treatment consists of up to 12 cycles.

  In one embodiment, the anti-PD-1 and anti-LAG-3 antibodies are administered as a first-line ("front-line") treatment (eg, a first or first treatment). In another embodiment, the anti-PD-1 and anti-LAG-3 antibodies use the same or different therapeutic agents, including second line treatments (eg, after relapse and / or when the first treatment fails). After the initial treatment). The anti-LAG-3 and anti-PD-1 antibodies can be administered to a subject by any suitable means. In one embodiment, the antibodies are formulated for intravenous administration. In another embodiment, the antibodies are administered simultaneously (eg, are formulated together in a single formulation, or concurrently with another formulation). Alternatively, in another embodiment, the antibodies are administered sequentially (eg, as separate formulations). In another embodiment, the anti-LAG-3 antibody is administered about 30 minutes (eg, about 29, 28, 27, 26, 25, 24, 23, 22, 21, 20 minutes or less) prior to administration of the anti-PD-1 antibody. ) Administered within.

  The efficacy of the treatment methods provided herein can be evaluated using any suitable means. In one embodiment, the treatment results in at least one therapeutic effect selected from the group consisting of reduced tumor size, reduced number of metastatic lesions over time, complete response, partial response, and stable disease.

Also, a therapeutically effective amount of an anti-LAG-3 antibody such as BMS-986016 and an anti-PD-1 antibody such as BMS-936558 and a pharmaceutically acceptable amount adapted for use in the methods described herein. Also provided is a kit comprising a pharmaceutical composition containing the carrier to be performed. In one embodiment, the kit comprises
(A) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 5 Anti-LAG-3 antibody;
(B) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 21 An anti-PD-1 antibody; and (c) instructions for using the anti-LAG-3 and anti-PD-1 antibodies in the methods of the invention.

  In another embodiment, in at least one cycle, the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR2 of the light chain variable region having the sequence set forth in SEQ ID NO: 21. CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3, and light chain variable having the sequence set forth in SEQ ID NO: 5, for co-administration with an anti-PD-1 antibody comprising a CDR3 domain Anti-LAG-3 antibody comprising the CDR1, CDR2 and CDR3 domains of the region, wherein for each cycle, 4 doses of anti-LAG-3 antibody are administered at a dose of 3, 20, 80 or 240 mg, and 4 doses of anti-PD Anti-LAG-3 antibody is provided wherein the -1 antibody is administered at a dose of 80 or 240 mgIn another embodiment, 4 doses of the anti-LAG-3 antibody are administered at a dose of 0.03, 0.25, 1 or 3 mg / kg body weight, and 4 doses of the anti-PD-1 antibody are 1 or 3 mg / kg. Administered in body weight doses.

  In another aspect of the invention, the anti-PD-1 antibody in any of the above embodiments is replaced by or combined with an anti-PD-L1 or anti-PD-L2 antibody. Accordingly, the present invention also provides for the treatment of tumors in human patients using said clinically effective dose of anti-LAG-3 antibody in combination with said clinically effective dose of anti-PD-1 antibody. Methods, compositions and kits, wherein the dose of the PD-1 antibody is replaced by the same dose of the anti-PD-L1 or anti-PD-L2 antibody.

FIG. 1 shows the inhibition of tumor growth in vivo using a combination treatment of anti-LAG-3 and anti-PD-1 antibodies in a mouse tumor model. 2A and 2B are schematic diagrams illustrating portions of a Phase I clinical trial. 2A and 2B are schematic diagrams illustrating portions of a Phase I clinical trial. FIG. 3 is a schematic diagram illustrating the screening, treatment, clinical follow-up, and survival follow-up phases of a clinical trial.

I. Definitions As used herein, the term "subject" or "patient" is a human cancer patient (e.g., a patient with an advanced solid tumor, such as a refractory solid tumor).

  As used herein, “effective treatment” refers to a treatment that results in a beneficial effect, eg, amelioration of at least one symptom of a disease or disorder. The beneficial effect may take the form of an improvement compared to a baseline, ie, an improvement compared to a measurement or observation made prior to initiation of therapy by the method. The beneficial effect may also take the form of stopping, slowing down, delaying, or stabilizing the detrimental progression of a marker of a solid tumor. Effective treatment may refer to alleviation of at least one symptom of a solid tumor. Such an effective treatment may, for example, reduce pain in the patient, reduce the size and / or number of lesions, reduce or prevent tumor metastasis, and / or slow tumor growth. .

  The term "effective amount" refers to an amount of an agent that provides a desired biological, therapeutic, and / or prophylactic result. The result is a reduction, amelioration, alleviation, reduction, delay, and / or alleviation of one or more of the signs, symptoms, or causes of the disease, or any other desired change in the biological system. Is also good. With reference to solid tumors, an effective amount will cause tumor shrinkage and / or reduce the rate of tumor growth (such as inhibiting tumor growth), or prevent or delay other undesirable cell growth. Contain sufficient amount to allow In some embodiments, the effective amount is an amount sufficient to delay tumor development. In some embodiments, the effective amount is an amount sufficient to prevent or delay tumor recurrence. An effective amount can be administered in one or more administrations. An effective amount of a drug or composition will (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) partially inhibit, delay, or slow cancer cell invasion into peripheral organs. (Iv) inhibits metastasis, ie, slows and stops to some extent; (v) inhibits tumor growth; (vi) prevents the appearance and / or recurrence of tumors, or Delay; and / or (vii) alleviate one or more symptoms associated with the cancer. In one example, an “effective amount” is the amount of anti-LAG-3 antibody and the amount of anti-LAG-3 antibody clinically proven to affect a significant reduction or slowing of cancer progression, such as in advanced solid tumors. Combination with the amount of PD-1 antibody. As used herein, the terms "fixed dose", "uniform dose" and "uniform fixed dose" are used interchangeably and refer to a dose administered to a patient independent of the patient's body weight or body surface area (BSA). . A fixed or flat dose is therefore provided not as a mg / kg dose, but rather as an absolute amount of drug (eg, anti-LAG-3 and / or anti-PD-1 antibody).

  As used herein, a “body surface area (BSA) based dose” is a dose that is adjusted to the body surface area (BSA) of an individual patient (eg, anti-LAG-3 antibody and / or anti-PD). -1 antibody). A dose based on BSA can be provided as mg / kg body weight. Various calculations have been published to reach BSA without direct measurement, the most widely used being the Du Bois equation (Du Bois D, Du Bois EF (Jun 1916) Archives of Internal Medicine 17 (6) ): 863-71; and Verbraecken, J. et al. (Apr 2006). Metabolism-Clinical and Experimental 55 (4): 515-24). Other exemplary BSA formulas include the Mosteller formula (Mosteller RD. N Engl J Med., 1987; 317: 1098) and the Haycock formula (Haycock GB, et al., J Pediatr 1978, 93: 62-66). ), Gehan and George's formula (Gehan EA, George SL, Cancer Chemother Rep 1970, 54: 225-235), Boyd's formula (Current, JD (1998), The Internet Journal of Anesthesiology 2 (2); and Boyd, Edith (1935), University of Minnesota. The Institute of Child Welfare, Monograph Series, No. x. London: Oxford University Press), Fujimoto's formula (Fujimoto S, et al., Nippon Eiseigaku Zasshi 1968; 5: 443-50) ), Takahira formula (Fujimoto S, et al., Nippon Eiseigaku Zasshi 1968; 5: 443-50), and Schlich formula (Schlich E, et al., Ernaehrungs Umschau 2010; 57: 178-183). .

  The term “antibody” describes a polypeptide that includes at least one antigen-binding site derived from an antibody (eg, a VH / VL region or Fv, or CDR). Antibodies include known forms of antibodies. For example, the antibody may be a human, humanized, bispecific, or chimeric antibody. The antibody may also be a Fab, Fab'2, ScFv, SMIP, Affibody (R), Nanobody, or domain antibody. Antibodies can also be of any of the following isotypes: IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgAsec, IgD, and IgE. Antibodies can be naturally occurring or altered (eg, by mutation, deletion, substitution, conjugation to a non-antibody portion). For example, an antibody may include one or more amino acid variants (as compared to a native antibody) that alter the properties (eg, functional properties) of the antibody. For example, some such changes are known in the art and affect, for example, the half-life of the antibody, effector function, and / or immune response in the patient. The term antibody also includes engineered polypeptide constructs that include at least one antibody-derived antigen binding site.

  The term "LAG-3" refers to lymphocyte activating gene-3. The term "LAG-3" includes variants, isoforms, homologs, orthologs and paralogs. For example, antibodies specific for human LAG-3 protein can, in certain cases, cross-react with LAG-3 protein from non-human species. In other embodiments, the antibody specific for the human LAG-3 protein may be completely specific for the human LAG-3 protein and does not show species or other types of cross-reactivity, Cross-reacts with LAG-3 from certain other species but may not cross-react with LAG-3 from all other species (eg, cross-reacts with monkey LAG-3) But does not cross-react with mouse LAG-3). The term "human LAG-3" refers to a human LAG-3 sequence, such as the complete amino acid sequence of human LAG-3 having Genbank accession number NP_002277 (SEQ ID NO: 13). The term "mouse LAG-3" refers to a mouse LAG-3 sequence, such as the complete amino acid sequence of mouse LAG-3 having Genbank accession number NP_032505. LAG-3 is also known in the art, for example, as CD223. The human LAG-3 sequence may differ from human LAG-3 at Genbank Accession No. NP_002277, for example, by having conserved mutations or mutations in non-conserved regions, wherein LAG-3 is Genbank Accession No. NP_002277. Has substantially the same biological function as human LAG-3. For example, the biological function of human LAG-3 is to have an epitope in the extracellular domain of LAG-3 that is specifically bound by the antibodies of the present disclosure, or the biological function of human LAG-3 The function is to bind to MHC class II molecules.

  The term "monkey LAG-3" is intended to encompass LAG-3 proteins expressed by old and new world monkeys, such as, but not limited to, cynomolgus LAG-3 and rhesus monkey LAG-3. You. A representative amino acid sequence of monkey LAG-3 is the rhesus monkey LAG-3 amino acid sequence also deposited as Genbank accession number XM_001108923. Another representative amino acid sequence of monkey LAG-3 is the alternative rhesus monkey sequence of clone pa23-5 described in US2011 / 0150892A1. This alternative rhesus monkey sequence shows a single amino acid difference at position 419 compared to the sequence deposited in Genbank.

  Certain human LAG-3 sequences are generally at least 90% identical in amino acid sequence to human LAG-3 of Genbank accession number NP_002277, and as compared to LAG-3 amino acid sequences of other species (eg, mouse), Contains an amino acid residue that identifies the amino acid sequence. In certain cases, human LAG-3 may be at least 95%, or even at least 96%, 97%, 98%, or 99% identical in amino acid sequence to LAG-3 of Genbank accession number NP_002277. In certain embodiments, the human LAG-3 sequence exhibits no more than 10 amino acid differences from the LAG-3 sequence at Genbank accession number NP_002277. In certain embodiments, human LAG-3 exhibits no more than 5, or even no more than 4, 3, 2, or 1 amino acid difference from the LAG-3 sequence of Genbank accession number NP_002277. Is also good. Percent identity can be determined as described herein.

  The terms "programmed death 1", "programmed cell death 1", "protein PD-1", "PD-1", "PD1", "PDCD1", "hPD-1" and "hPD-" used herein. “1” is used interchangeably and includes variants, isoforms, species homologs, and analogs of PD-1 that have at least one common epitope with PD-1. The complete PD-1 sequence can be found in GenBank Accession No. U64863 (SEQ ID NO: 29).

  The programmed death 1 (PD-1) protein is an inhibitory member of the CD28 family of receptors and also includes CD28, CTLA-4, ICOS and BTLA. PD-1 is expressed on activated B cells, T cells, and myeloid cells (Agata et al., Supra; Okazaki et al. (2002) Curr. Opin. Immunol. 14: 391779-82; Bennett et. al. (2003) J Immunol 170: 711-8). Early members of this family, CD28 and ICOS, were discovered by a functional effect on increasing T cell proliferation following addition of monoclonal antibodies (Hutloff et al. Nature (1999); 397: 263-266; Hansen et al. Immunogenics (1980); 10: 247-260). PD-1 was discovered by screening for differential expression in apoptotic cells (Ishida et al. EMBO J (1992); 11: 3887-95). Other members of the family, CTLA-4 and BTLA, were discovered by screening for differential expression in cytotoxic T lymphocytes and TH1 cells, respectively. CD28, ICOS and CTLA-4 all have unpaired cysteine residues that allow homodimerization. In contrast, PD-1 is suggested to exist as a monomer lacking the characteristic unpaired cysteine residue in other CD28 family members.

  The PD-1 gene is a 55 kDa type I transmembrane protein that is part of the Ig gene superfamily (Agata et al. (1996) Int Immunol 8: 765-72). PD-1 contains a membrane proximal immune receptor tyrosine inhibitory motif (ITIM) and a membrane distal tyrosine-based switch motif (ITSM) (Thomas, ML (1995) J Exp Med 181: 1953-6; Vivier, E and Daeron, M (1997) Immunol Today 18: 286-91). Although structurally similar to CTLA-4, PD-1 lacks the MYPPPY motif, which is important for B7-1 and B7-2 binding. Two ligands for PD-1 have been identified, PD-L1 and PD-L2, which have been shown to down-regulate T cell activation upon binding to PD-1 (Freeman et al. (2000) | Exp Med 192: 1027-34; Latchman et al. (2001) Nat Immunol 2: 261-8; Carter et al. (2002) Eur J Immunol 32: 634-43). PD-L1 and PD-L2 are both B7 homologs that bind to PD-1, but do not bind to other CD28 family members. PD-L1 is abundant in various human cancers (Dong et al. (2002) Nat. Med. 8: 787-9). Interaction between PD-1 and PD-L1 results in reduced tumor infiltrating lymphocytes, reduced T cell receptor-mediated proliferation, and immune evasion by cancerous cells (Dong et al. (2003)). Mol. Med. 81: 281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54: 307-314; Konishi et al. (2004) Clin. Cancer Res. 10: 5094-100). Immunosuppression can be reversed by inhibiting the local interaction between PD-1 and PD-L1, the effect being similar when the interaction between PD-1 and PD-L2 is blocked. Nat'l. Acad. Sci. USA 99: 12293-7; Brown et al. (2003) J. Immunol. 170: 1257-66.

Consistent with PD-1, an inhibitory member of the CD28 family, PD-1 deficient animals develop a variety of autoimmune phenotypes, including autoimmune cardiomyopathy and lupus-like syndrome with arthritis and nephritis (Nishimura et al. (1999) Immunity 11: 141-51; Nishimura et al. (2001) Science 291: 319-22). In addition, PD-1 has been found to play a role in autoimmune encephalomyelitis, systemic lupus erythematosus, graft-versus-host disease (GVHD), type I diabetes, and rheumatoid arthritis (Salama et al. (2003) J Exp Med 198: 71-78; Prokunina and Alarcon-Riquelme (2004) Hum Mol Genet 13: R143; Nielsen et al. (2004) Lupus 13: 510). In a mouse B cell tumor line, the ITSM of PD-1 has been shown to be essential for blocking BCR-mediated Ca2 ⁺ flux and tyrosine phosphorylation of downstream effector molecules (Okazaki et al. (2001). ) PNAS 98: 13866-71).

  "Programmed death ligand-1 (PD-L1)" is one of two cell surface glycoprotein ligands for PD-1 that down-regulates T cell activation and cytokine secretion upon binding to PD-1 (The other is PD-L2). As used herein, the term "PD-L1" refers to human PD-L1 (hPD-L1), variants, isoforms and species homologs of hPD-L1, and at least one common with hPD-L1. Includes 5 analogs with epitopes. The complete hPD-L1 sequence can be found under GenBank accession number Q9NZQ7.

IIa. Anti-LAG-3 Antibodies Anti-human LAG-3 antibodies (or VH / VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, art-recognized anti-LAG-3 antibodies can be used. For example, using the anti-human LAG-3 antibody described in US2011 / 0150892A1, referred to as monoclonal antibody 25F7 (also known as "25F7" and "LAG3.1"), the teachings of which are incorporated herein by reference. can do. Other art-recognized anti-LAG-3 antibodies that can be used include IMP731 described in US2011 / 007023, the teachings of which are also incorporated herein by reference.

  Antibodies that compete with any of the art-recognized antibodies referenced above for binding to LAG-3 can also be used.

  An exemplary anti-LAG-3 antibody is a heavy chain comprising the sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively, described in PCT / US13 / 48999, the teachings of which are incorporated herein by reference. BMS-986016 comprising a light chain, or antigen-binding fragments and variants thereof.

  In another embodiment, the antibody has the heavy and light chain CDRs or variable region of BMS-986016. Thus, in one embodiment, the antibody comprises a CDR1, CDR2, and CDR3 domain of the VH region of BMS-986016 having the sequence set forth in SEQ ID NO: 3, and a VL region of BMS-986016 having the sequence set forth in SEQ ID NO: 5. CDR1, CDR2 and CDR3 domains. In another embodiment, the antibody is a CDR1, CDR2 and CDR3 domain comprising the sequence set forth in SEQ ID NOs: 7, 8 and 9, respectively, and a CDR1, CDR2 and CDR2 comprising the sequence set forth in SEQ ID NOs: 10, 11 and 12, respectively. Includes CDR3 domain. In another embodiment, the antibody comprises a VH and / or VL region comprising the amino acid sequence set forth in SEQ ID NO: 3 and / or SEQ ID NO: 5, respectively. In another embodiment, the antibody comprises the variable heavy (VH) and / or variable light (VL) regions encoded by the nucleic acid sequences set forth in SEQ ID NO: 4 and / or SEQ ID NO: 6, respectively. In another embodiment, the antibody competes for and / or binds to the same epitope on LAG-3 as the antibody described above. In another embodiment, the antibody binds to an epitope of human LAG-3 comprising the amino acid sequence PGHPLAPG (SEQ ID NO: 14). In another embodiment, the antibody binds to an epitope of human LAG-3 comprising the amino acid sequence HPAAPSSSW (SEQ ID NO: 15) or PAAPSSWG (SEQ ID NO: 16).

  In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity with an antibody described above (eg, at least about 90%, 95% or 99% variable amino acid identity with SEQ ID NO: 3 or SEQ ID NO: 5). Region identity).

IIb. Anti-PD-1 Antibodies Suitable anti-human PD-1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, art-recognized anti-PD-1 antibodies can be used. For example, monoclonal antibody 5C4 (referred to herein as nivolumab or BMS-936558), 17D8, 2D3, 4H1, 4A11, 7D3, described in WO 2006/121168, the teachings of which are incorporated herein by reference. And 5F4 can be used. Other known PD-1 antibodies include lambrolizumab (MK-3475) as described in WO2008 / 156712 and AMP-514 as described in WO2012 / 145493, the teachings of which are incorporated herein by reference. . Additional known PD-1 antibodies and other PD-1 inhibitors include those described in WO2009 / 014708, WO03 / 099996, WO2009 / 114335 and WO2011 / 161699, the teachings of which are incorporated herein by reference. Things. Antibodies that compete with any of these art-recognized antibodies or inhibitors for binding to PD-1 can also be used.

  An exemplary anti-PD-1 antibody is BMS-936558 comprising a heavy and light chain comprising the sequences set forth in SEQ ID NO: 17 and SEQ ID NO: 18, respectively, or an antigen binding fragment and variant thereof.

  In another embodiment, the antibody has the heavy and light chain CDRs or variable region of BMS-936558. Thus, in one embodiment, the antibody is a CDR1, CDR2 and CDR3 of VH of BMS-936558 having the sequence set forth in SEQ ID NO: 19 and a CDR1, CDR2 of VL of BMS-936558 having the sequence set forth in SEQ ID NO: 21. And CDR3. In another embodiment, the antibody is a CDR1, CDR2, and CDR3 domain comprising the sequence set forth in SEQ ID NOS: 23, 24, and 25, respectively, and a CDR1, CDR2, and CDR2 comprising the sequence set forth in SEQ ID NOs: 26, 27, and 28, respectively. Includes CDR3 domain. In another embodiment, the antibody comprises a VH and / or VL region comprising the amino acid sequence set forth in SEQ ID NO: 19 and / or SEQ ID NO: 21, respectively. In another embodiment, the antibody comprises a variable heavy (VH) and / or variable light (VL) region encoded by the nucleic acid sequence set forth in SEQ ID NO: 20 and / or SEQ ID NO: 22, respectively. In another embodiment, the antibody competes for and / or binds to the same epitope on PD-1 as the antibody described above. In another embodiment, the antibody has at least about 90% variable region amino acid sequence identity to an antibody described above (eg, at least about 90%, 95% or 99% variable amino acid identity to SEQ ID NO: 19 or SEQ ID NO: 21). Region identity).

IIc. Anti-PD-L1 Antibodies Suitable anti-human PD-L1 antibodies (or VH and / or VL domains derived therefrom) suitable for use in the present invention can be produced using methods well known in the art. Alternatively, art-recognized anti-PD-L1 antibodies can be used. For example, the human anti-PD-L1 antibody disclosed in U.S. Patent No. 7,943,743, the contents of which are incorporated herein by reference, can be used. Such anti-PD-L1 antibodies include 3G10, 12A4 (also referred to as BMS-936559), 10A5, 5F8, 10H10, 1B12, 7H1, 11E6, 12B7, and 13G4. Other art-recognized anti-PD-L1 antibodies that can be used include, for example, U.S. Patent Nos. 7,635,757 and 8,217,149, U.S. Patent Application Publication No. 2009/0317368. And those described in PCT Publication Nos. WO2011 / 066389 and WO2012 / 145493, the teachings of which are also incorporated herein by reference. Antibodies that compete with any of these art-recognized antibodies or inhibitors for binding to PD-L1 can also be used.

III. Pharmaceutical Compositions Pharmaceutical compositions suitable for administration to human patients are typically formulated for parenteral administration, e.g., in a liquid carrier, or liquid solution for intravenous administration or Suitable for reconstitution in suspension.

  Generally, such compositions will typically include a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" is approved by a governmental regulatory authority or is another generally recognized for use in the United States Pharmacopeia or animals, particularly humans Meaning listed in Pharmacopoeia. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such pharmaceutical carriers are also sterile liquids, such as water and oils, including those of petroleum, animal, plant or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, glycerol polyethylene glycol silinolic acid, and the like. Good. Water or aqueous solutions saline and aqueous dextrose and glycerol solutions can be employed as carriers, especially for injectable solutions (including, for example, anti-LAG-3 or anti-PD-1 antibodies). Liquid compositions for parenteral administration can be formulated for administration by injection or continuous infusion. Routes of administration by injection or infusion include intravenous, intraperitoneal, intramuscular, intrathecal and subcutaneous. In one embodiment, the anti-LAG-3 and / or anti-PD-1 antibodies are administered intravenously [eg, in separate formulations or together (in the same or separate formulations)].

IV. Patient Populations Provided herein are methods for treating solid tumor cancers (eg, advanced refractory solid tumors) in human patients using a combination of an anti-LAG-3 antibody and an anti-PD-1 antibody. This is a clinical method.

  Examples of cancers that can be treated using the methods of the present invention include liver cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, breast cancer, lung cancer, skin cancer Or malignant melanoma in the eye, kidney cancer, uterine cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, cancer in the anal area, stomach cancer, testicular 20 cancer, uterine cancer, egg Ductal carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, non-Hodgkin's lymphoma, esophagus cancer, small intestine cancer, endocrine system cancer, thyroid cancer, Cancer of the parathyroid gland, adrenal gland, soft tissue sarcoma, urethra cancer, penis cancer, solid tumor in childhood 25, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, Renal pelvis cancer, central nervous system (CNS) neoplasm, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma Environment-induced cancers such as pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, and those induced by asbestos, eg, multiple myeloma, B-cell lymphoma, Hodgkin's lymphoma / primary mediastinum B Cell lymphoma, non-Hodgkin's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, immunoblastic large cell lymphoma, progenitor Hematologic malignancies 30, such as B lymphoblastic lymphoma, mantle cell lymphoma, acute lymphoblastic leukemia, mycosis fungoides, anaplastic large cell lymphoma, T cell lymphoma, and precursor T lymphoblastic lymphoma, and Any combination of the above cancers is included. The present invention is also applicable to the treatment of metastatic cancer.

  In one embodiment, the human patient has non-small cell lung cancer (NSCLC) or a virus-related cancer (eg, a human papillomavirus (HPV) -related tumor) or gastric adenocarcinoma. In certain embodiments, the HPV-related tumor is HPV + head and neck cancer (HNC). In another specific embodiment, the gastric adenocarcinoma is associated with an Epstein-Barr virus (EBV) infection.

  Patients can be tested or selected for one or more of the above clinical attributes before, during, or after treatment.

V. Combination Therapy The combination therapy provided herein comprises an anti-LAG-3 antibody and an inhibitory immunoreceptor (eg, its natural receptor) for treating a subject having a solid tumor (eg, an advanced refractory solid tumor). Including the administration of another antibody, particularly an anti-PD-1 antibody, which blocks a receptor that inhibits / neutralizes activities such as cytotoxic activity upon binding to a ligand.

  In one embodiment, the present invention provides a combination of an anti-LAG-3 antibody and an anti-PD-1 antibody with a defined clinical dosing regimen for treating a subject having a solid tumor (eg, a refractory solid tumor). I will provide a. In certain embodiments, the anti-LAG-3 antibody is BMS-986016. In another embodiment, the anti-PD-1 antibody is BMS-936558. In another embodiment, the dosing regimen is adjusted to provide the optimal desired dose (eg, an effective response).

  As used herein, additive or combined administration (co-administration) includes simultaneous administration of the compounds in the same or different dosage forms, or separate administration of the compounds (eg, sequential administration). Thus, the anti-LAG-3 and anti-PD-1 antibodies can be administered simultaneously in a single formulation. Alternatively, the anti-LAG-3 antibody and the anti-PD-1 antibody can be formulated for separate administration and administered simultaneously or sequentially (eg, one antibody is administered as a second antibody). Administered within about 30 minutes prior to antibody administration).

  For example, the anti-LAG-3 antibody can be administered after the anti-PD1 antibody is first administered (eg, immediately after), or vice versa. In one embodiment, the anti-PD-1 antibody is administered prior to the administration of the anti-LAG-3 antibody. In another embodiment, the anti-PD-1 antibody is administered after administration of the anti-LAG-3 antibody. In another embodiment, the anti-LAG-3 and anti-PD-1 antibodies are administered simultaneously. By such simultaneous or sequential administration, preferably both antibodies are present simultaneously in the treated patient.

VI. Treatment Protocols Suitable treatment protocols for treating solid tumors in human patients include, for example,
(A) an anti-LAG- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 5; 3 antibodies,
(B) an anti-PD- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 19, and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 21; Administering to the patient an effective amount of each of the 1 antibodies,
The method includes at least one dosing cycle, the cycle being eight weeks, wherein at least four doses of the anti-LAG-3 antibody are provided in each of at least one cycle of about 1, 3, 10, 20, 50, 80, 100, 130. , 150, 180, 200, 240 or 280 mg in a uniform dose, and at least 4 doses of the anti-PD-1 antibody in about 50, 80, 100, 130, 150, 180, 200, 240 or 280 mg in a uniform dose Is done. In another embodiment, the four doses of anti-LAG-3 antibody are 0.01, 0.03, 0.25, 0.1, 0.3, 1 or 3, 5, 8, or 10 mg / kg body weight. The dose is administered and 4 doses of the anti-PD-1 antibody are administered at a dose of 0.1, 0.3, 1, 3, 5, 8, or 10 mg / kg body weight.

In one embodiment, the anti-LAG-3 and anti-PD-1 antibodies are:
(A) 3 mg of anti-LAG-3 antibody and 80 mg of anti-PD-1 antibody;
(B) 3 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(C) 20 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(D) 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(E) Administered at a dose of 240 mg anti-LAG-3 antibody and 240 mg anti-PD-1 antibody.

In another embodiment, the anti-LAG-3 and PD-1 antibodies are
(A) 0.3 mg / kg anti-LAG-3 antibody and 1 mg / kg anti-PD-1 antibody;
(B) 0.3 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody;
(C) 0.25 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody;
(D) 1 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody; or (e) administered at a dose of 3 mg / kg anti-LAG-3 antibody and 3 mg / kg anti-PD-1 antibody Is done.

  In one embodiment, the dose of the anti-LAG-3 and / or anti-PD-1 antibody is calculated per body weight, for example, in mg / kg body weight. In another embodiment, the dose of the anti-LAG-3 antibody and / or anti-PD-1 antibody is a fixed fixed dose. In another embodiment, the dose of the anti-LAG-3 and / or anti-PD-1 antibodies varies over time. For example, anti-LAG-3 and / or anti-PD-1 antibodies can be initially administered at high doses and decreased over time. In another embodiment, the anti-LAG-3 and / or anti-PD-1 antibodies are initially administered at low doses and are increased over time.

  In another embodiment, the amount of anti-LAG-3 and / or anti-PD-1 antibody administered is constant for each dose. In another embodiment, the amount of antibody administered varies with each dose. For example, the maintenance (or continuation) dose of the antibody may be higher than or the same as the initially administered loading dose. In another embodiment, the maintenance dose of the antibody may be lower than or the same as the loading dose.

  In another embodiment, the anti-LAG-3 and / or anti-PD-1 antibodies are formulated for intravenous administration. In one embodiment, the anti-PD-1 antibody is administered on days 1, 15, 29 and 43 of each cycle. In another embodiment, the anti-LAG-3 antibody is administered on days 1, 15, 29 and 43 of each cycle.

  In other embodiments, the anti-LAG-3 and / or anti-PD-1 antibodies have weekly, biweekly or triweekly, monthly, or clinical benefit observed. Administered as long as possible or until there is a complete response, confirmed progressive disease or unmanageable toxicity.

  In another embodiment, the cycle of administration is 8 weeks and can be repeated as needed. In another embodiment, the treatment consists of up to 12 cycles.

  In another embodiment, four doses of the anti-PD-1 antibody are administered per eight week cycle. In another embodiment, four doses of an anti-LAG-3 antibody are administered per eight week cycle.

  In another embodiment, the anti-PD-1 and anti-LAG-3 antibodies are administered as a first-line treatment (eg, an initial or first-line treatment). In another embodiment, the anti-PD-1 and anti-LAG-3 antibodies are used in a second-line treatment (eg, after an initial or first treatment, such as after relapse and / or when the first treatment fails). Later).

  In another aspect, the invention features any of the above embodiments, wherein the anti-PD-1 antibody is replaced by or combined with an anti-PD-L1 or anti-PD-L2 antibody.

VII. Outcome For the target lesion, response to therapy may include:

  For non-target lesions, response to therapy may include:

  Preferably, patients treated according to the methods disclosed herein will experience improvement in at least one symptom of cancer. In one embodiment, the improvement is measured by a decrease in the amount and / or size of a measurable tumor lesion. In another embodiment, lesions can be measured on chest x-ray or CT or MRI film. In another embodiment, cytology or histology can be used to assess responsiveness to therapy.

  In one embodiment, the patient to be treated is a complete response (CR), a partial response (PR), a stable disease (SD), an immune-related complete disease (irCR), an immune-related partial response (irPR), or an immune response. 2 shows a related stable disease (irSD). In another embodiment, the patient to be treated experiences tumor shrinkage and / or reduced growth rate, ie, suppression of tumor growth. In another embodiment, unwanted cell growth is reduced or inhibited. In yet another embodiment, one or more of the following can be performed: the number of cancer cells can be reduced; tumor size can be reduced; cancer cell invasion into peripheral organs Inhibit, delay, slow down, or arrest; can slow down or inhibit tumor metastasis; can inhibit tumor growth; prevent or delay tumor recurrence Be able to alleviate one or more symptoms associated with cancer.

  In another embodiment, administration of an effective amount of an anti-LAG-3 antibody and an anti-PD-1 antibody by any of the methods provided herein reduces tumor size, the number of metastatic lesions that appear over time, At least one therapeutic effect selected from the group consisting of reduced, complete remission, partial remission, or stable disease. In yet other embodiments, the method of treatment results in a substantial clinical benefit (CBR = CR + PR + SD ≧ 6 months) better than that achieved with only anti-LAG-3 or anti-PD-1 antibodies. In other embodiments, the improvement in clinical margin is about 20%, 30%, 40%, 50%, 60%, 70%, 80% compared to the anti-LAG-3 or PD-1 antibody alone. % Or more.

VIII. Kits and Unit Dosage Forms Also, a therapeutically effective amount of an anti-LAG-3 antibody, such as BMS-986016, and an anti-PD-1 antibody, such as BMS-936558, in a therapeutically effective amount adapted for use in the method. A kit comprising a pharmaceutical composition comprising an acceptable carrier is also provided. The kit also allows a practitioner (eg, a physician, nurse, or patient) to administer the composition contained in the kit to a patient having a cancer (eg, a solid tumor). And, where appropriate, for example, instructions including a dosing schedule. The kit may also include a syringe.

  The kit may comprise a plurality of packages of a single dose of a pharmaceutical composition each containing an effective amount of an anti-LAG-3 or anti-PD-1 antibody for a single administration according to the methods provided above. The equipment or devices necessary to administer the pharmaceutical composition can also be included in the kit. For example, a kit can provide one or more pre-filled syringes containing an amount of an anti-LAG-3 or anti-PD-1 antibody.

In one embodiment, the present invention provides a kit for treating a solid tumor in a human patient,
(A) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 5 Anti-LAG-3 antibody;
(B) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 21 An anti-PD-1 antibody; and (c) said kit comprising instructions for using the anti-LAG-3 and anti-PD-1 antibodies in the methods described herein.

  The following examples are merely illustrative, and should not be construed as limiting the scope of the disclosure in any way, as many modifications and equivalents will become apparent to those skilled in the art upon reading the present disclosure. Absent.

  The contents of all references, GenBank registrations, patents and published patent applications cited throughout this application are expressly incorporated herein by reference.

Preclinical pharmacology of anti-PD-1 antibody (BMS-936558) BMS-936558 has a nanomolar affinity and high specificity as measured by surface plasmon resonance using a Biacore® biosensor system. 1 is a fully human IgG4 (kappa) isotype monoclonal antibody that binds to C.1, thus making it impossible to bind to its ligands PD-L1 and PD-L2. BMS-936558 does not bind to other related family members, such as BTLA, CTLA-4, ICOS or CD28. Preclinical studies of BMS-936558 have shown that binding to PD-1 results in enhanced T cell proliferation and release of interferon-gamma (IFN-gamma) in vitro. The heavy and light chain amino acid sequences of BMS-936558 are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

In vivo toxicity of the anti-PD-1 antibody novolumab (BMS-936558) Toxicity studies in cynomolgus monkeys show that BMS-936558 is well tolerated in doses up to 50 mg / kg administered twice weekly, up to 27 doses It was confirmed that. Drug-related findings were limited to a 28% reversible decrease in triiodothyronine (T3), with no abnormalities in other markers of thyroid function (data not shown).

Clinical pharmacology and safety of anti-PD-1 antibody (BMS-936558) The overall safety experience with BMS-936558 as a monotherapy or in combination with other therapeutic agents has been reported to date with approximately 1500 patients treated to date. It is based on experience in human subjects. In general, for monotherapy, the safety profiles are similar between tumor types. One exception is pulmonary infiltration adverse events (AEs), which, in some cases, make it difficult to distinguish pulmonary symptoms and radiographic changes from those associated with BMS-936558. Since there may be, the numbers may be higher in subjects with NSCLC. Safety profiles were generally consistent across completed and ongoing clinical trials, with none of the doses up to 10 mg / kg tested reaching maximum tolerated doses. There was no pattern in the incidence, severity, or cause of adverse events at the BMS-936558 dose level.

  Study CA209003 has contributed a great deal of clinical experience with BMS-936558 to date in subjects with NSCLC and other solid malignancies. CA209003 was a phase I multiple dose escalation study in subjects with previously treated advanced or metastatic melanoma, RCC, NSCLC, colorectal cancer, or hormone refractory prostate cancer. In CA209003, subjects received 0.1, 0.3, 1, 3, or 10 mg / kg of BMS-936558 intravenously every two weeks. No maximum tolerance was identified in CA209003. The maximum dose level evaluated was 10 mg / kg. The incidence, severity, and relationship of adverse events were generally similar across dose levels and tumor types.

  As of July 3, 2012, at least one adverse event was reported, regardless of cause, in 296 (97.4%) of the 304 subjects treated with BMS-936558. There was no pattern in the incidence, severity, or relationship of adverse events to BMS-936558 dose levels. Either grade of treatment-related adverse events occurred in 220 (72.4%) subjects. The most frequent drug-related adverse events that occur in more than 5% of subjects are fatigue (25.7%), rash (13.5%), diarrhea (11.8%), pruritus (10.2%), nausea ( 7.9%), decreased appetite (7.9%), decreased hemoglobin (5.9%), and fever (5.3%). Many treatment-related adverse events were of low grade (grade 1 or 2). Treatment-related high-grade (grade 3 or 4) adverse events were reported in 45 subjects (14.8%), the most common being fatigue (1.6%) and decreased appetite (1.0%) , And diarrhea (1.0%). At all dose levels, at least one serious adverse event (SAE) was reported in 150 of the 304 subjects (49.3%). Grade 3-4 SAEs were reported for 23 subjects (7.6%). Either grade of drug-related SAE occurred in 11.5% of subjects. Grade 3-4 drug-related SAEs reported in at least two subjects include diarrhea (3 subjects [1.0%]), pneumonitis (3 subjects [1.0%]), pneumonia ( 2 subjects [0.7%]), and lipase elevation (2 subjects [0.7%]). As with the overall adverse event profile, there was no apparent relationship in the incidence or severity of drug-related adverse events to BMS-936558 dose levels. There were no apparent differences in the frequency of adverse events based on the tumor type of the subject. Selected treatment-related adverse events occurred infrequently (less than 5%) but are considered clinically relevant because they require higher alertness for early recognition and immediate intervention. These adverse events included elevated alanine aminotransferase (ALT) (4.3%), elevated aspartate aminotransferase (AST) (3.6%), pneumonia (3.3%), and decreased thyroid function (3%). 2.0%), hyperthyroidism (1.3%), adrenal dysfunction (0.7%), and colitis (0.7%). Pneumonitis grade 3-4 events were reported in three subjects (1.0%) as described above (one event was grade 4). Grade 3 events of colitis, ALT elevation, and AST elevation were reported in two subjects (0.7%) each. Grade 3 events of adrenal dysfunction, hyperthyroidism, and hypothyroidism were each reported in one subject (0.3%). Due to possible clinically relevant BMS-936558-related adverse events requiring early recognition and immediate intervention, due to suspected pulmonary toxicity, suspected diarrhea or colitis, hepatotoxicity, endocrine disorders, and nephrotoxicity A management algorithm was developed.

  Treatment-related adverse events leading to withdrawal were reported in 18 of the 304 treated subjects (5.9%) on CA209003. The only events reported in more than one subject were pneumonitis (4 subjects [1.3%]) and hepatitis (2 subjects [0.7%]). There were three (1.0%) drug-related deaths; each occurred after the onset of pneumonitis. The safety of BMS-936558 in combination with other therapeutic agents has been investigated in several ongoing clinical trials.

Pharmacokinetics of Anti-PD-1 Antibody (BMS-936558) The pharmacokinetics (PK) of a single dose of BMS-936558 was determined for 39 patients with multiple tumor types in CA209001 at a dose range of 0.3-10 mg / kg. It was evaluated in the subject. The median Tmax over the dose levels ranged from 1.6 to 3.1 hours, with individual values ranging from 0.9 to 7 hours. The PK of BMS-936558 is linear in the range of 0.3 to 10 mg / kg, with a dose proportional increase in Cmax and AUC (INF) and low to moderate inter-subject variability observed at each dose level. (I.e., the coefficient of variation [CV] ranged from 16-45%). The geometric mean clearance (CLT) after a single IV dose ranged from 0.13 to 0.19 mL / h / kg, but the mean volume of distribution (Vz) varied between doses from 83 to 113 mL / kg. The mean terminal T-HALF of BMS-936558 was 17-25 days, consistent with the half-life of endogenous IgG4, indicating that the excretion mechanism of BMS-936558 may be similar to that of IgG4. Is shown. Both elimination and distribution of BMS-936558 were considered to be dose independent within the dose range tested. In available data from 128 subjects in a multiple dose study of multiple tumor types (CA209003), the mean T-HALF was 21-24 hours and the median T-max was 0.6-3 across dose levels. 0.3, consistent with the single dose data.

Phase I Clinical Trial for Anti-PD-1 Antibody (BMS-936558) BMS-936558 is one completed Phase 1 single dose study and ongoing in subjects with NSCLC, melanoma, RCC, and other malignancies Showed clinical activity in two multiple dose escalation studies (Phase 1 monotherapy: Phase 1b combination therapy with CA209003 and ipilimumab). Tumor response was determined by the Response Evaluation Criteria in Modified Solid Tumors (RECIST) established by the NCI. The evaluable population consists of 294 subjects with various solid malignancies (melanoma, n = 138; NSCLC, n = 122; RCC, n = 34) currently treated with nivolumab.

  In CA209003, an objective response rate (ORR) of 31.1% (33 out of 106 response evaluable subjects) was observed with BMS-936558 monotherapy at doses ranging from 0.1 to 10 mg / kg. Reported in subjects with melanoma treated every two weeks (Q2W). Most responses were persistent, exceeding 6 months.

  In many active dose ranges (3-10 mg / kg), ORR is 13.5% to 27.8% in subjects with NSCLC and progression-free survival (PFSR) at 24 weeks is 23% to 51%. It was reported that there was. Sustained responses were observed in both flat and non-flat subtypes.

  Of the 34 response evaluable RCC subjects in CA209003, 1 mg / kg (5 of 18 subjects, 27.8%) and 10 mg / kg (5 of 16 subjects, 31. 3%) Responses were reported in both treatment groups. Progression-free survival (PFSR) estimates at 24 weeks were 50% in the 1 mg / kg BMS-936558-treated group and 67% in the 10 mg / kg BMS-936558-treated group.

  Preliminary results from a Phase 1b study of the combination therapy of BMS-936558 with ipilimumab suggest the benefit of combining two T cell targeted therapies for subjects with melanoma. In the 0.3 mg / kg BMS-936558 + 3 mg / kg ipilimumab treatment group, responses were observed in 5 out of 14 evaluable subjects (35.7%, conventional modified World Health Organization [mWHO]. One complete response and two partial responses by criteria, and two partial responses by immune-related mWHO criteria). In the 1 mg / kg BMS-936558 + 3 mg / kg ipilimumab treatment group, responses were observed in 9 out of 15 evaluable subjects (60%, 3 CRs and 6 PRs; all conventional mWHO standard). In the 3 mg / kg BMS-936558 + 3 mg / kg ipilimumab treatment group, an objective response was observed in 4 out of 6 evaluable subjects (66.7%, 3 parts by conventional mWHO criteria) Response and one partial response according to immune-related mWHO criteria). Further details are provided in Wolchok et al. (2013) NEJM 369 (2): 122-33, and / or PCT / US2013 / 040767.

Preclinical Pharmacology of Anti-LAG-3 Antibody (BMS-986016) BMS-986016 is a fully human antibody specific for human LAG-3 isolated from immunized transgenic mice expressing the human immunoglobulin gene. BMS-986016 is expressed as an IgG4 isotype antibody, which is a stabilized hinge that attenuates Fc receptor binding to reduce or eliminate the possibility of antibody or complement mediated killing of target cells. Includes mutation (S228P). The heavy and light chain amino acid sequences of BMS-986016 are set forth in SEQ ID NO: 17 and SEQ ID NO: 18, respectively.

The ability of BMS-986016 to bind to recombinant human LAG-3 antigen was determined using Biacore and an enzyme-linked immunosorbent assay (ELISA). Binding to human and primate LAG-3 + transfectants and activated human or primate T cells was measured using flow cytometry and Scatchard analysis. BMS-986016 binds human LAG-3 with high affinity (K _D = 0.12-0.5 nM) and inhibits binding of LAG-3 to cells expressing its ligand, MHC class II (IC50, 0.67 nM). BMS-986016 has a lower affinity for cynomolgus LAG-3 on transfected CHO cells and activated cynomolgus T cells than on activated human T cells (EC50, 21.5-34.3 nM). ). In the absence of a second costimulator, high concentrations of BMS-986016 do not elicit a measurable cytokine response from cultured human peripheral blood cells, and the drug has no measurable antibody-dependent or It does not mediate complement-dependent killing. BMS-986016 promotes the activation of antigen-specific mouse T cell hybridomas expressing human LAG-3 in co-culture with MHC class II positive antigen presenting cells. In addition, BMS-986016, when added alone or in combination with BMS-936558 (anti-PD-1 antibody), enhances activation of human T cells in a superantigen stimulation assay.

Toxicity of anti-LAG-3 antibody (BMS-986016) alone or in combination with anti-PD-1 antibody (BMS-936558) The following preclinical toxicity studies were performed:
A. Exploratory Combination Pharmacodynamics and Toxicity Studies by Intermittent Intravenous (QW) 4-Week Administration in Cynomolgus Monkeys Using Anti-LAG3.1 Antibody (Precursor to BMS-986016) and BMS-936558. Was. Anti-LAG3.1 administered alone or in combination with 50 mg / kg / week of BMS-936558 at 50 mg / kg / week did not produce any deleterious changes. The single-agent anti-toxic effect level [No-observed-adversese-effect level (NOAEL)] for anti-LAG3.1 is 50 mg / kg / week (AUC [0-168h] = 231,000 μg · h / mL). NOAEL for anti-LAG3.1 in combination with 50 mg / kg / week of BMS-936558 was 50 mg / kg / week (mean anti-LAG3.1 AUC [0-168h] = 210,000 μg · h / mL; mean BMS-936558 AUC [0-168h] = 159,500 μg · h / mL).

B. GLP-Based Combinatorial Toxicity Test by Intravenous Administration for 4 Weeks in Cynomolgus Monkeys with 6 Weeks Recovery Using BMS-986016 and BMS-936558 Important results were as follows. Single agent BMS-986016, administered at a maximum of 100 mg / kg / week, did not cause any adverse changes. Single agent BMS-936558, administered at 50 mg / kg / week, results in minimal to minimal irreversible lymphoplasmic cell inflammation of the choroid plexus of the brain, which is a combination with BMS-986016 and BMS-936558 Considered to be non-harmful considering the lower severity and incidence of lymphoplasma inflammation compared to treatment, lack of vasculitis or tissue destruction, and the absence of clinical signs during the course of treatment Was done. Combination administration of BMS-986016 and BMS-936558 (100 and 50 mg / kg / week, respectively) resulted in the mortality of one male out of nine monkeys on day 29 of the study. From day 26 to day 29, the monkeys had increased body temperature, tremors, red or clear nasal discharge, altered feces (amorphous feces, insufficient feces, or absence of feces), reduced feeding behavior, mild She exhibited dehydration, sneezing, hypoactivity, and a stooped posture. Two days after veterinary care and antibiotic treatment, the animals showed no improvement and were euthanized on day 29 due to poor clinical condition.

  There were no significant gross necropsy findings. Histopathological findings in this monkey include slight lymphoplasmic inflammation of the choroid plexus; minimal to moderate lymphohistiocytic inflammation of the vasculature of the brain parenchyma, meninges, and spinal cord (cervical and lumbar); It contained minimal to moderate mixed cell inflammation of the epididymis, seminal vesicles and testes. Clinicopathological changes showed an unexplained decrease in red blood cell count, hemoglobin concentration and hematocrit, and increased fibrinogen correlated with inflammation observed in the central nervous system (CNS) and male reproductive organs.

  Further histopathological findings upon the combined administration of BMS-986016 and BMS-936558 (100 and 50 mg / kg / week, respectively) indicate that minimal choroid plexus of the brain in 7 of the remaining 8 monkeys. Minimal to minimal irreversible lymphoplasmic cell inflammation and minimal lymphohistiocytic inflammation of the vasculature of the brain parenchyma in one of the remaining eight monkeys (the reversibility of which cannot be assessed Was limited to).

  The NOAEL for single agent BMS-986016 was considered to be 100 mg / kg / week (mean AUC [0-168h] = 474,000 μg · h / mL); the NOAEL for single agent BMS-936558 was 50 mg / kg. / Week (mean AUC [0-168h] = 193,000 μg · h / mL); the NOAEL for the combination of BMS-986016 and BMS-936558 was not determined. However, the combination therapy was generally well tolerated and clinical signs of toxicity were observed in only one of the nine monkeys (about 10%). Thus, 100/50 mg / kg / week of BMS-986016 / nivolumab (mean BMS-986016 AUC [0-168h] = 514,000 μg · h / mL; mean nivolumab AUC [0-168h] = 182,000 μg · h / mL) was considered STD10.

  The doses administered (100 mg / kg BMS-986016 and 50 mg / kg BMS-936558) are more than 10-fold higher than the maximum dose proposed for this study. The starting dose of BMS-986016 monotherapy (Part A), 20 mg (0.25 mg / kg), is less than 1/10 of the human equivalent of cynomolgus NOAEL (636 mg; 8.0 mg / kg), 265 times Less than HED (24 mg; 0.30 mg / kg) after linear adjustment of NOAEL target exposure for estimated maximum affinity difference. The safety multiple calculated for exposure at the starting dose of 20 mg (0.25 mg / kg) is 100 mg / kg / week for the cynomolgus monkey NOAEL, without accounting for affinity differences. Is 315 times.

  The starting dose of 3 mg (0.03 mg / kg) of BMS-986016 in the combination therapy (Part B) is a linear adjustment of the cynomolgus monkey STD10 for a maximum affinity difference estimated at 265-fold, plus a 10-fold safety factor based on. The maximum recommended starting dose (MRSD) for BMS-986016 based on 100 mg / kg / week STD10 is 0.03 mg / kg in humans. The starting dose of BMS-936558 in the combination therapy (Part B) of 80 mg (1 mg / kg) is based on the PK parameters of known human BMS-936558 plus a 10-fold safety factor. The MRSD of BMS-936558 based on cyno STD10 at 50 mg / kg / week was 4.3 mg / kg in humans and was further reduced to identify doses showing acceptable levels of adverse events.

C. GLP-Based Tissue Cross-Reactivity Tests in Human and Selected Cynomolgus Monkey Tissues Using BMS-986016 Positive staining with BMS-986016-FITC was observed in the following human tissues: bladder, blood cells, colon-colon, eye Mononuclear leukocytes of the esophagus, small intestine, stomach, kidney, lung, lymph node, placenta, salivary gland, skin, spleen, thymus, tonsils, cervix and endometrium; and cell membranes or cell membrane granules of hematopoietic cells of the bone marrow . In addition, staining with BMS-986016-FITC was observed in the cytoplasm of human pituitary secretory cell epithelium. Within a limited panel of cynomolgus tissues evaluated, staining with BMS-986016-FITC was observed in the plasma membrane or plasma membrane granules of spleen mononuclear leukocytes. LAG-3 expressing cells with lymphocyte morphology and distribution in the germinal center and interfollicular T cell regions of normal human lymphoid tissues (lymph node, tonsils, spleen, thymus, bone marrow and mucosa-associated lymphoid tissues) Scientific reports predicted the staining of mononuclear leukocytes and hematopoietic cells by BMS-986016-FITC in this study (in human and cynomolgus monkey tissues). Considering that LAG-3 mRNA was expressed in human pituitary and LAG3.1-G4P-FITC staining was observed in the anterior pituitary gland of human pituitary in a pilot tissue cross-reactivity study, BMS-986016-FITC staining of cell epithelial cytoplasm and cytoplasmic granules was also expected. Although BMS-986016 is not expected to have access to the cytoplasmic compartment in vivo and repeated dose toxicity studies in monkeys showed no effect on the pituitary gland, these findings may be clinically relevant .

D. Evaluation of in vitro cytokine release and lymphocyte activation for BMS-986016 using human peripheral blood mononuclear cells. Was not induced. The observed levels of cytokines were at or near the lower assay limit of quantification, with no evidence of dose-dependence or pattern between donors (IL-1β, IL-2, IL-5, IL-10 , IL-12p70, and IFN-γ), or, generally, overlapped with cytokine levels from PBMC incubated with negative controls (IL-6, IL-8, TNF-α).

  Consistent with the lack of cytokine release, there was no evidence that BMS-986016 induced T or NK cell activation as measured by CD25 and CD69 surface expression. The expression levels of these markers on T and NK cells after stimulation with BMS-986016 were similar to those observed upon stimulation with a negative control.

  Collectively, these data indicate that BMS-986016 has no agonistic ability to induce T or NK cell activation or cytokine release.

Preclinical pharmacokinetics of anti-LAG-3 antibody (BMS-986016) Regulatory guidelines for biotechnology-derived pharmaceuticals (ICH Harmonized Tripartite Guideline, S6 (R1) Preclinical Safety Evaluation of Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceuticals, Pharmaceutical Sciences, Pharmaceutical Sciences, Pharmaceuticals, Pharmaceuticals, Pharmaceutical Sciences, Pharmaceutical Sciences, Pharmaceutical Sciences, Biotechnology, USA) Metabolic studies on BMS-986016 were not performed in animals. The expected in vivo degradation of monoclonal antibodies (mAbs) is to small peptides and amino acids by biochemical pathways that are independent of the cytochrome P450 enzymes.

  BMS-986016 showed favorable pharmacokinetic (PK) properties in cynomolgus monkeys. From both single dose and repeated dose IV PK studies, BMS-986016 degraded biexponentially and exposure was approximately dose proportional. Whole body clearance (CLTp) was 0.12-0.22 mL / h / kg, and terminal half-life (T-HALF) was 133-414 hours. The steady state distribution volume (Vss) was 62-72 mL / kg, suggesting that the extraplasma distribution is limited. Although anti-BMS-986016 antibody was detected in some monkeys, the presence of anti-BMS-986016 antibody did not appear to have an effect on BMS-986016 exposure.

Inhibition of tumor growth in vivo by combined treatment of anti-LAG-3 and anti-PD-1 antibodies To test the hypothesis that the combination of anti-LAG-3 and anti-PD-1 antibodies enhances antitumor efficacy Next, an experiment was performed in a mouse tumor model. These studies evaluated tumor growth inhibition in syngeneic tumor models (Sa1N fibrosarcoma and MC38 colon adenocarcinoma) and monitored enhanced autoimmunity in non-obese diabetes (NOD) models. Administration of anti-LAG-3 antibody resulted in both total tumor growth inhibition and an increase in the number of tumor-free (TF) mice in these treatment groups, as shown in FIG. Anti-LAG-3 antibody administered in combination with anti-PD-1 antibody resulted in enhanced anti-tumor activity beyond that of either drug alone. For example, in multiple Sa1N tumor models, 20% -30% TF mice with anti-LAG-3 antibody compared to control and anti-PD-1 antibody treated mice (0% -10% TF mice) Was obtained, but 60% to 90% of TF mice were obtained by combining the anti-LAG-3 antibody and the anti-PD-1 antibody. In the MC38 model, the anti-LAG-3 antibody showed only modest tumor growth inhibition, but when administered in combination with the anti-PD-1 antibody, the enhancement was greater than that observed for the anti-PD-1 antibody alone Antitumor activity was obtained (80% vs. 40% TF mice, respectively).

Phase 1 Study in Patients with Solid Tumors A phase 1 study of anti-LAG-3 antibody (BMS-986016) and anti-PD-1 antibody (BMS-936558) administers BMS-986016 and BMS-936558 as a combination treatment. Performed in patients with advanced solid tumors to prove efficacy.

1. Objectives The primary objective of this study was to evaluate the safety and tolerability of BMS-986016 administered in combination with BMS-936558 in subjects with advanced solid tumors, as well as dose limiting toxicity (DLT) and maximal It is to identify the tolerance (MTD).

  The second objective was to evaluate the preliminary antitumor activity of the combination of BMS-986016 and BMS-936558 in subjects with advanced solid tumors, the pharmacokinetics of BMS-986016 and BMS-936558 when co-administered ( PK), monitoring the immunogenicity of BMS-986016 and BMS-936558 administered as a combination therapy, and the effects of BMS-986016 and BMS-936558 on corrected QT ("QTc") Including evaluating. A further exploratory objective was to evaluate the pharmacodynamic effects of BMS-98616 and BMS-936558 combination therapy based on selected biomarkers in peripheral blood and tumor biopsy specimens, and to assess the pharmacodynamic effects of BMS-98616 and BMS-936558. Characterizing T cell function during combination therapy, assessing 2-year landmark overall survival in subjects treated with BMS-986016 and BMS-936558, BMS-986016 and BMS in subjects with advanced solid tumors Exploring the preliminary anti-tumor activity of the combination therapy with 936558, characterizing the pharmacokinetic and exposure response relationships in subjects treated with BMS-986016 and BMS-936558, and selecting peripheral and selected clinical efficacy To examine the relationship with tumor biomarkers Including the.

2. Study Design and Duration This is a phase 1, open-label study of BMS-986016, administered as a single agent and in combination with BMS-936558 (nivolumab) to subjects with advanced solid tumors. The test is performed in three parts. Part A and Part B consist of a 3 + 3 + 3 dose escalation design using BMS-986016 administered alone (Part A) or in combination with BMS-936558 (Part B) in subjects with advanced solid tumors. Treatment in Part B begins with the decision to escalate to the third dose cohort in Part A (follows the dose escalation rule). Subsequently, the recruitment in the two parts proceeds simultaneously. There is no point where the dose of BMS-986016 in combination with BMS-936558 (Part B) exceeds the dose of BMS-986016 previously shown to be safe on the monotherapy dose escalation group (Part A). Part C consists of a cohort expansion in a six disease limited population of approximately 16 subjects each using BMS-98558 administered in combination with BMS-936558. The treatment in Part C is initiated when the maximum tolerated dose (MTD) for Part B (or if the MTD is not determined, the maximum dose (MAD)). The dose selected for Part C does not exceed the MTD or MAD of Part B, but the titration may include evaluation of toxicity and other data, including PK and pharmacodynamic data from Parts A and B. A schematic of the test is shown in FIG.

  Subjects complete the study for up to four periods as follows: screening (up to 28 days), treatment (up to 12 8-week cycles of test therapy), clinical follow-up (135 days), and survival follow-up ( Up to two years after the first dose of study drug; longer efficacy follow-up periods may be considered in selected cases if the efficacy signal is evident). During this period, diagnostic imaging can be performed every 12 weeks until progression in subjects discontinued due to CR and subjects with PR at the end of cycle 12.

  The treatment period consists of up to 12 8-week treatment cycles. Each treatment cycle consisted of 4 doses of BMS-986016 alone (Part A) or BMS-986016 in combination with BMS-936558 (Parts B and C) administered on days 1, 15, 29 and 43 of each treatment cycle. ). In Parts B and C, where both antibodies are administered in combination, nivolumab is administered first, then BMS-986016 within 30 minutes of completing the infusion of nivolumab. Tumor response is assessed using RECIST v1.1. Subjects may have (1) complete response (CR) confirmation, (2) completion of a maximum of 12 8-week cycles, (3) progressive disease (PD), (4) clinical deterioration, and / or 5) Continue study therapy until any of the other criteria for discontinuation is met. Certain patients with PD defined by the initial RECIST v1.1 who have received clinical benefit and are being treated will continue to be treated after progression. Subjects who discontinue treatment enter a 135-day clinical follow-up period.

  After the completion of the clinical follow-up period, the subject enters the survival follow-up period. During this period, a clinic visit or telephone contact is performed every 12 weeks to assess survival. The duration of this period can be up to two years after the first administration of the study drug, but longer follow-up periods are possible in certain cases if an efficacy signal is evident. Subjects in the survival follow-up with advanced disease will receive tumor therapy as needed. A schematic of the test is set forth in FIG.

  Evaluations, including physical examination, vital sign measurements, 12-lead ECG, and laboratory evaluation, are performed at selected time points throughout the dosing interval. Subjects will be closely monitored for adverse events throughout the study. Blood samples are taken for pharmacokinetic analysis up to 4 hours after the start of study drug administration.

  Subjects will continue therapy for up to 12 eight-week cycles or until confirmation of complete response, progressive disease, clinical deterioration, or fulfillment of discontinuation criteria. Subjects received a total of 28 days of screening, up to 12 8-week cycles of treatment, 135 days of clinical follow-up, and up to 2 years of follow-up (starting with the first dose of study drug). For up to about 2.3 years. The total duration of the study is expected to be approximately 5 years from the first visit of the first subject to the required survival follow-up of the last subject enrolled.

3. Dose escalation Part A
In Part A, the safety of BMS-986016 administered as a single agent is evaluated using a 3 + 3 + 3 design. If the subject can begin the first day of dosing within about one week of a third subject in the same dose escalation cohort, a fourth subject can be enrolled at the start of the dose escalation cohort. Dose escalation doses are shown in FIGS. 2A and 2B and Table 1 (described below). Three subjects (or four, as appropriate) are treated first in each dose cohort. In dose cohort 1, each of the first three subjects (or four, if appropriate) is designated as a monitor and treatment is initiated at least every five days. Subjects in subsequent cohorts do not need to be observed at 5 day intervals on the day of treatment initiation. The dose escalation in Part A proceeds as follows:
-If 0 of the first 3 subjects (or 4 if appropriate) experience dose limiting toxicity (DLT) within the DLT evaluation interval, 3 subjects (or 4 if appropriate) ) Will be treated at the next higher dose level.
If one of the three subjects (or four if appropriate) experiences DLT within the DLT assessment interval, extend the cohort to six subjects.
If two of the six subjects experience DLT within the DLT assessment interval, extend that cohort to nine subjects.
• 2 or more of 3 (or 4 if appropriate) subjects, 3 or more of 6 subjects, or 3 or more of 9 subjects dosed during DLT evaluation interval If DLT is experienced within the cohort, the dose level is determined to be above the MTD.

  Any cohort previously established as safe before expanding to MTD (or MAD) may be expanded to gain further experience or to achieve dose levels intermediate to those defined in the protocol. consider. Dose escalation rules (cohort size, DLT evaluation interval, cohort expansion criteria, etc.) also apply to these expanded or additional cohorts. A maximum of 9 subjects will be enrolled in any additional or extended dose cohort.

  Intra-subject dose escalation is not acceptable. If a dose level is found to be above the MTD, subjects enrolled at that dose level will be treated at a lower dose.

Part B
Treatment in Part B begins after the decision to escalate to the third dose cohort is made in Part A (following the dose escalation rule). Subsequently, the recruitment in the two parts proceeds simultaneously. At any time point, the dose of BMS-986016 administered in combination with BMS-936558 (Part B) is the dose of BMS-986016 previously shown to be safe in the monotherapy dose escalation group (Part A) Never exceed. The assignment of treatments to subjects that qualify for both Part A and Part B is alternating between the two parts, and subjects to be treated subsequently via the Interactive Voice Response System (IVRS) whenever possible Assigned to different parts. If there is no opening available in the part to which the object is assigned by this algorithm, the object is assigned to the next open cohort or part. As in Part A, the 3 + 3 + 3 design is also used in Part B to evaluate the safety of BMS-986016 administered in combination with nivolumab. If the first day of dosing can begin within about one week of a third subject in the same dose escalation cohort, a fourth subject can be enrolled at the start of the dose escalation cohort. The dose evaluated during dose escalation is shown in FIGS. 2A and 2B and Table B (described below). As in Part A, each of the first three subjects (or four, if appropriate) in the first dose cohort in Part B are designated as monitored and treatment is initiated at least every five days.

  Three subjects will be treated first in each dose cohort. In dose cohort 1, each of the first three subjects designated as monitored will begin treatment at least every 5 days. Subjects in subsequent cohorts do not need to be maintained at 5 day intervals between treatment initiation dates.

  The dose escalation in Part B proceeds as described for Part A. If the MTD is exceeded in dose cohort 2, subsequent cohorts will be treated with 20 mg BMS-986016 and 80 mg BMS-936558. If this dose combination is found to be safe, escalation will proceed with the previously defined BMS-986016 dose, maintaining the BMS-936558 dose at 80 mg.

  If the MTD is not reached through dose cohort 5, an additional cohort of BMS-986016 administered in combination with BMS-936558 will be considered based on the overall safety experience during dose escalation.

  Any cohort previously established as safe before expanding to MTD (or MAD) may be expanded to gain further experience or to achieve dose levels intermediate to those defined in the protocol. consider. Dose escalation rules (cohort size, DLT evaluation interval, cohort expansion criteria, etc.) also apply to these expanded or additional cohorts. Up to nine subjects will be enrolled in any boost or extended dose cohort.

  Intra-subject dose escalation is not acceptable. If a dose level is found to be above the MTD, subjects enrolled at that dose level are reduced to lower doses.

4. Cohort Expansion The purpose of the cohort expansion is to accumulate additional safety, tolerability, preliminary efficacy, pharmacokinetic, and pharmacodynamic information for the combination of BMS-986016 and BMS-936558. The dose selected for Part C does not exceed the MTD (or MAD if MTD is not determined) in Part B, but allows for the evaluation of other data, including toxicity and PK and pharmacodynamic data from Parts A and B. May be included. Doses include intermediate doses at the dose levels evaluated in Part B. If a dose below the MTD is selected, use modeling as an aid to inform about the choice of combination dose level to use in Part C. The six expanded cohorts are limited to the tumor types listed in Tables 3A and 3B below. A continuous assessment of toxic events in the cohort extension will be performed through enrollment in the extension cohort. At any time, further enrollment is discontinued if the integrated rate of treatment-related toxicity meeting the DLT criteria is greater than 33% across all subjects treated in the Part C cohort extension. Depending on the nature and grade of toxicity and after assessing the risk: benefit ratio, new doses for the entire cohort may be given at lower dose levels previously tested, or intermediate to lower dose levels previously tested. Start with.

  Once the MTD in Part A (or MAD if MTD is not determined), the BMS-986016 monotherapy cohort is evaluated in cohort expansion. This expanded cohort is limited to tumor types that have been found to be responsive to BMS-986016 monotherapy. The dose selected for monotherapy expansion should not exceed the MTD of Part A (or MAD if MTD is not determined), and be based on other data including toxicity and PK and pharmacodynamic data from Part A. Consider the evaluation. The dose chosen is intermediate between the dose levels tested in Part A. If a dose below the MTD is selected, use modeling as an aid to inform about the choice of dose level to use in Part C.

5. Dose Limiting Toxicity BMS-986016 has the potential to increase the frequency and severity of previously described adverse events associated with BMS-936558, or to introduce new toxicity. To guide decisions regarding dose escalation in Part A and Part B, harm associated with study drug and occurring within 56 days (8 weeks) of study drug initiation (ie, DLT assessment interval to completion of cycle 1) Dose limiting toxicity (DLT) is determined based on the incidence, intensity, and duration of the event. Adverse event severity is graded according to the National Cancer Institute (NCI) Common Terminology Criteria for Advance Events (CTCAE) v4.0. If DLT occurred at any time, whether during dose escalation (Part A and Part B) or cohort expansion (Part C), for subject management, all studies The drug is stopped and the assessment of the event's relevance to the test drug is suspended. To restart the test procedure, the subject must first meet the criteria for re-treatment.

  Subjects withdrawing from the study during the DLT evaluation interval for reasons other than DLT can be replaced with the same dose level. If an infusion cannot be administered at the scheduled visit during the DLT evaluation interval, it must be administered as soon as possible. If the delay is 1-7 days, the original scheduled visit procedure should be performed and the subject is considered evaluable for DLT determination. If the delay is more than 7 days, it is considered not administered and is not replaced. Subjects made 3 of 4 scheduled BMS-986016 doses in Part A (or 4 scheduled in Part B) throughout the 8-week observation period to make decisions regarding dose escalation from safety predictions (3 times of BMS-986016 and nivolumab doses), a subject is considered evaluable only if that one dose that did not result in a progressive disease or for non-medical reasons. Non-evaluable subjects can be replaced at the same dose level.

  Liver, non-hematological, and hematological DLTs are defined separately as outlined below.

Liver DLT
Any of the following drug-related events are considered liver DLT:
ALT or AST> 8 x ULN, regardless of duration, or cannot return to grade 1 or less within 2 weeks despite medical intervention, ALT or AST> 5 x and ≤ 8 x ULN, or Total bilirubin> 5 × ULN, or ALT or AST> 3 × ULN and simultaneous total bilirubin> 2 × ULN.

Non-hematological DLT
Any of the following drug-related events are considered nonhematological DLTs:
Grade 2 immune-related ocular pain or vision loss requiring systemic treatment, or Or non-liver or non-hematological toxicity of grade 3 or higher, except those described below.

The following grade 3 non-hematological events are not considered DLTs:
Grade 3 electrolyte abnormalities lasting less than 72 hours, not clinically deteriorating, and resolving spontaneously, or responding to conventional medical intervention. Persistent for less than 3 weeks, and clinical or x-ray of pancreatitis. Grade 3 increase in amylase or lipase not associated with image evidence lasting less than 48 hours, and resolved spontaneously or by conventional medical intervention to grade 1 or lower, grade 3 nausea or vomiting lasting less than 72 hours Grade 3 endocrine disorders that are well controlled by grade 3 fever / hormone replacement, and are not associated with poor hemodynamics (including clinical or laboratory evidence of hypotension or end-organ perfusion injury) Grade 3 tumor flare (defined as pain, irritation, or rash localized at the site of a known or suspected tumor)
-Grade 3 fatigue of less than 7 days.

Hematological DLT
Any of the following drug-related events is considered a hematological DLT:
Grade 4 febrile neutropenia of any duration. Grade 4 neutropenia lasting longer than 5 days. Grade 4 thrombocytopenia. Grade 4 anemia. Clinically significant bleeding. Grade 3 thrombocytopenia associated with: Grade 3 febrile neutropenia lasting longer than 48 hours. Grade 3 hemolysis.

6. Study Patient Criteria Signed Informed Consent Subjects must sign an IRB / IEC-approved informed consent document prior to performing any study-related procedures that are not considered part of standard treatment. Must be dated.

  Consent for Tumor Biopsy Samples for Part A or Part B Dose Escalation: Subjects will be present on block or unstained slides of existing formalin-fixed paraffin-embedded (FFPE) tumor tissue for the purpose of conducting correlation studies. You must agree to allow any acquisition. If archived samples are not available, subjects must agree to allow tumor biopsy before treatment. In each case, the investigator must ensure that the tissue block or slide is physically present before starting therapy. Subjects who cannot provide archived tumor samples, disagree with pre-treatment tumor biopsies, or have no harvestable lesions are ineligible (however, the amount of tissue obtained by pre-treatment biopsy or Objects of poor quality are not ineligible on this basis.)

  Consent for Tumor Biopsy Samples for Part C Cohort Expansion: Subjects with Melanoma or Head and Neck Tumors: All Subjects in Two Melanoma Cohorts and All Subjects in Head and Neck Tumor Cohorts It is necessary to undergo a test; therefore, the subject must have a lesion at a location that can be determined by the investigator to be able to obtain the specimen at an acceptable clinical risk. The biopsy site for any subject must be different from the evaluable lesion. Subjects in the melanoma and head and neck cancer cohorts that do not meet these criteria are ineligible; however, subjects with insufficient tissue quality or quality from screening biopsies are not ineligible on this basis . Subjects in the remaining cohort (NSCLC or gastric adenocarcinoma): Subjects agree to allow the acquisition of blocked or unstained slides of existing formalin-fixed paraffin-embedded (FFPE) tumor tissue for the purpose of conducting correlation studies. There must be. If archived samples are not available, subjects must agree to allow tumor biopsy before treatment. In each case, the investigator must ensure that the tissue block or slide is physically present before starting therapy. Subjects who cannot provide archived tumor samples, disagree with pre-treatment tumor biopsies, or have no harvestable lesions are ineligible (however, the amount of tissue obtained by pre-treatment biopsy or Objects of poor quality are not ineligible on this basis.) Even if available, biopsies cannot be taken in subjects with only a single measurable lesion.

Target Population a) Subjects must have histological or cytological confirmation of advanced (metastatic and / or unresectable) non-curable solid malignancy:
i) Part A. Dose escalation: BMS-986016 monotherapy (1) All histological solid tumor cases except subjects with primary CNS tumors are acceptable;
(2) Without limitation, CTLA-4, ipilimumab, tremelimumab, anti-PD-1, anti-PD-L1, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibody, etc. Only subjects who have not been previously exposed to the immune cell regulatory antibody regimen (ICMAR) are allowed;
(3) If the subject has received at least one standard treatment regimen in an advanced or metastatic setting, if such therapy is present, unless the subject has progressed or is not tolerated by the standard treatment regimen No.

ii) Part B. Dose escalation: BMS-986016 + BMS-936558
(1) All histological solid tumor cases except those with primary CNS tumors are acceptable. Subjects with or without prior anti-PD-1 or anti-PD-L1 antibody therapy are eligible. Alternatively, except for subjects having a primary CNS tumor, but are not limited to, anti-CTLA-4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-PD-L2 antibody, anti-KIR antibody, anti-KIR antibody, All histological solid tumor cases naive to ICMAR, such as the CD137 antibody or the anti-OX40 antibody, are accepted.
(2) Subjects without prior anti-PD-1 antibody therapy or anti-PD-L1 antibody therapy include, but are not limited to, ipilimumab, tremelimumab, anti-PD-L2 antibody, anti-KIR antibody, anti-CD137 antibody Or any other ICMAR, such as anti-OX40 antibodies, must not have been previously exposed. Alternatively, subjects without prior anti-PD-1 or anti-PD-L1 antibody therapy include, but are not limited to, ipilimumab, tremelimumab, anti-PD-L2 antibody, anti-KIR antibody, anti-CD137 antibody, Or have not been previously exposed to any other ICMAR, such as an anti-OX40 antibody.
(3) NSCLC subjects whose disease progresses during or after therapy with anti-PD-1 or anti-PD-L1 antibody as the most recent therapy. Alternatively, the subject where the most recent therapy is a previous anti-PD-1 or anti-PD-L1 antibody:
(A) Disease is unresponsive during anti-PD-1 or anti-PD-L1 antibody therapy and presents with PD (by RECIST) within 16 weeks of therapy initiation.
(B) The therapy had been discontinued due to serious and / or life-threatening anti-PD-1 or anti-PD-L1 antibody-related toxicity (eg, dose limiting toxicity in previous studies). must not.
(C) Informed consent must be provided within 60 days after the last dose of anti-PD-1 or anti-PD-L1 antibody therapy.
(D) Previously exposed to ICMAR such as, but not limited to, anti-CTLA-4 antibody therapy, ipilimumab, tremelimumab, anti-PD-L2 antibody, anti-KIR antibody, anti-CD137 antibody, or anti-OX40 antibody. Don't wait.
(4) Melanoma subjects whose disease has progressed during or after receiving anti-CTLA-4 antibody therapy and anti-PD-1 antibody therapy or anti-PD-L1 antibody therapy. (A) Anti-CTLA-4 antibody therapy and anti-PD-1 or anti-PD-L1 antibody therapy may be received on a continuous or combination regimen. (B) The last dose of anti-CTLA-4 antibody therapy must have been received at least 100 days after the first dose of study drug. (C) Therapy must not have been discontinued due to serious and / or life-threatening antibody-related toxicity (eg, dose limiting toxicity in previous studies). (D) It must not have been previously exposed to any ICMAR other than anti-CTLA-4 antibody therapy and anti-PD-1 or anti-PD-L1 antibody therapy.
(5) Subjects must have progressed after receiving at least one standard treatment regimen, or have not been tolerated by that standard treatment regimen.

iii) Part C. Cohort expansion (1) The following groups will be registered:
(A) Melanoma-subject naive to ICMAR (b) Melanoma-disease is non-responsive during the most recent anti-PD-1 or anti-PD-L1 antibody therapy and within 16 weeks of starting therapy Subject with PD (according to RECIST). Because subjects must provide informed consent within 60 days after the last dose of anti-PD-1 or anti-PD-L1 antibody therapy, severe and / or life-threatening toxicity (eg, previous Antibody therapy should not have been discontinued due to dose limiting toxicity in the study). These subjects have previously been exposed to any other ICMAR such as, but not limited to, ipilimumab, tremelimumab, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibodies. Don't wait. Alternatively, subjects with disease progression while receiving (either in a continuous or combination regimen) receiving anti-CTLA-4 and anti-PD-1 or anti-PD-L1 antibody therapy are eligible. The last dose of anti-CTLA-4 antibody therapy must be received at least 100 days after the first dose of study drug. Subjects should not have discontinued antibody therapy due to severe and / or life-threatening toxicity (eg, dose limiting toxicity in previous studies). These subjects must not have been previously exposed to any ICMAR other than anti-CTLA-4 antibody therapy and anti-PD-1 or anti-PD-L1 antibody therapy.
(C) Non-small cell lung cancer (NSCLC)-a subject naive to ICMAR.
(D) NSCLC-Subjects who are non-responsive to disease during anti-PD-1 or anti-PD-L1 antibody therapy as the most recent therapy and exhibit PD (by RECIST) within 16 weeks of therapy initiation. Because subjects must provide informed consent within 60 days after the last dose of anti-PD-1 or anti-PD-L1 antibody therapy, severe and / or life-threatening toxicity (eg, previous Antibody therapy should not have been discontinued due to dose limiting toxicity in the study). These subjects have previously been exposed to any other ICMAR such as, but not limited to, ipilimumab, tremelimumab, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibodies. Don't wait. Alternatively, NSCLC subjects whose disease progresses during or after therapy using anti-PD-1 or anti-PD-L1 antibodies as the most recent therapy. Subjects should not have discontinued antibody therapy due to severe and / or life-threatening toxicity (eg, dose limiting toxicity in previous studies). These subjects must not have been previously exposed to any other ICMAR, such as anti-CTLA-4, anti-PD-L2, anti-KIR, anti-CD137, or anti-OX40 antibodies.
(E) HPV-related head and neck tumors-naive to ICMAR and HPV positive as defined by p16 immunohistochemistry (IHC) positive and / or HPV-16 in situ hybridization (ISH) positive Target. (I) Histological cases are limited to squamous cell carcinoma. Alternatively, subjects with advanced / metastatic head and neck tumors-naive to ICMAR. (I) Histological cases are limited to squamous cell carcinoma.
(F) Gastric adenocarcinoma subjects naive to ICMAR.
(I) HER2 (+) and HER2 (-) targets are allowed.

b) the subject must have undergone at least one standard treatment regimen in an advanced or metastatic setting, if such a therapy is present, or have progressed or has not been tolerated by that standard treatment regimen .
c) Subjects with any previous treatment regime are eligible. The following are not considered separate lines of treatment: adding compounds to an ongoing regimen, restarting the same regimen after a drug holiday, or switching from IV to oral therapy.
d) The presence of at least one lesion with a measurable disease as defined by the RECIST v1.1 criteria for response assessment. Subjects with lesions in previously irradiated areas as the only site of measurable disease are allowed to register if the lesions show a clear progression prior to the time of informed consent and can be accurately measured Is done.
e) 0 or 1 ECOG state.
f) Life expectancy of 12 weeks or more at the time of informed consent.
g) sufficient organ function as defined by:
i) Leukocytes (WBC) of 2000 / μL or more (stable without growth factors within 4 weeks of the first test drug administration)
ii) 1500 / μL or more of neutrophils (stable without growth factors within 4 weeks of the first test drug administration)
iii) platelets greater than 100 × 10 ³ / μL (infusion to achieve this level is not allowed within 2 weeks of the first study drug administration)
iv) hemoglobin of 8.5 g / dL or higher (transfusion to achieve this level is not acceptable within 2 weeks of the first study drug administration)
v) Creatinine of less than 1.5 × ULN or creatinine clearance of 40 mL / min or more (Cockcroft-Gault formula)
vi) ALT and AST below 3xULN
vii) Lipase and amylase less than 1.5xULN viii) Total bilirubin less than 1.5xULN (except subjects with Gilbert syndrome who must have normal direct bilirubin)
h) Stable to normal thyroid function or hormone replacement.
i) Ability to follow treatment, PK, and pharmacodynamic sampling and required test follow-up.
j) Re-enrollment of subjects: This trial allows for re-enrollment of subjects who discontinued the study as pre-treatment failures (ie, non-randomized / untreated subjects). If re-registered, subject must agree again.

Age and reproductive status a) Men and women aged 18 years or older at the time of informed consent.
b) Birth-capable women (WOCBP) must use contraceptive methods. Two forms of contraception are needed for teratogenicity test drugs and / or when there is not enough information to assess teratogenicity (no preclinical studies have been performed). One method must be highly effective (less than 1% failure rate when used consistently and correctly), and the second method may be highly effective. Specific contraceptive methods should be determined in consultation with the investigator. WOCBP must follow contraceptive instructions for a total of 24 weeks (30 days plus the time required to receive the study drug 5 half-life) after the last dose of study drug. Birth-capable women (WOCBP) are defined as any non-menopausal women who have experienced menarche and have not undergone contraceptive surgery (hysterectomy or bilateral ovariectomy). Menopause is defined as 12 months of amenorrhea in women over the age of 45 in the absence of other biological or physiological causes. In addition, women under 55 must have documented serum follicle stimulating hormone (FSH) levels above 40 mIU / mL to confirm menopause. Women treated with hormone replacement therapy (HRT) may have artificially suppressed FSH levels and may require a period of elimination to achieve physiological FSH levels. The duration of the ejection period is a function of the type of HRT used. The duration of the elimination period below is a suggested guideline and the investigator should use that judgment in checking serum FSH levels. If serum FSH levels exceed 40 mIU / ml at any time during the elimination period, the woman can be considered postmenopausal.
A minimum of one week for vaginal hormone products (rings, creams, gels), a minimum of four weeks for transdermal products, and a minimum of eight weeks for oral products.
c) The woman must be negative in serum or urine pregnancy test within 24 hours before the start of study drug (urine pregnancy test of total human villous gonadotropin (hCG) or its beta fraction with a minimum sensitivity of 25 IU / L).
d) Women must not be breastfeeding.
e) Men who are sexually active with WOCBP must use contraceptive methods. Two forms of contraception are needed for teratogenicity test drugs and / or when there is not enough information to assess teratogenicity (no preclinical studies have been performed). One method must be highly effective (less than 1% failure rate when used consistently and correctly), and the second method may be highly effective. Men who are sexually active with WOCBP must follow contraceptive instructions for a total of 33 weeks (90 days plus the time required to receive the study drug 5 half-life) after the last dose of study drug.

  Infertile women (ie, postmenopausal or undergoing contraceptive surgery); and men with permanent azoospermia (eg, bilateral orchiectomy) do not require contraception. Birth-capable women (WOCBP) are defined as women who have experienced menarche and have not undergone contraceptive surgery (hysterectomy or bilateral ovariectomy) or have not been postmenopausal. Menopause is clinically defined as 6 months of amenorrhea in women over the age of 45 in the absence of other biological or physiological causes. In addition, women under 55 must have documented serum follicle stimulating hormone (FSH) levels above 40 mIU / mL to confirm menopause.

7. Exclusion Criteria Target Disease Exception Subjects with a known or suspected CNS metastasis or CNS as the sole site of active disease are excluded with the following exceptions:
i) Subjects with controlled brain metastases are allowed to enroll. Controlled brain metastasis is defined as no radiographic progression for at least 4 weeks after irradiation and / or surgical treatment at the time of consent. Subjects must have been on steroids for at least two weeks prior to informed consent and must have no new or progressive neurological signs and symptoms.
ii) Subjects with signs or symptoms of brain metastases are not eligible unless brain metastases are excluded by computed tomography (CT) or magnetic resonance imaging (MRI).
Participation in previous clinical trials with BMS-936558, such as subjects in the comparison arm, where overall survival was listed as primary or secondary primary endpoint and did not complete analysis based on primary endpoint.

History and concurrent disease Appropriately treated basal cell or squamous cell skin cancer, localized prostate cancer, cervical or bladder intraepithelial neoplasia, or noninvasive ductal or lobular carcinoma in situ Subjects with previous malignancies, except for, are excluded. Subjects with a malignancy diagnosed more than two years ago (at the time of informed consent) who have been treated for therapeutic purposes, have no evidence of disease during the interval, and have a low risk of recurrence Subjects considered to be eligible.
Subjects with isolated vitiligo, resolved childhood asthma / atopy, controlled adrenal or hypopituitarism, and euthyroid patients with a history of Graves' disease (controlled hyperthyroidism Have a history of any active or known or suspected autoimmune disease (except thyroglobulin and thyroid peroxidase antibodies and thyroid-stimulating immunoglobulin prior to study drug administration) Target.
A known or underlying medical condition that may adversely affect the subject's ability to administer, follow, or accept the test drug, which is dangerous to the subject.
Requirements for daily oxygen supplementation.
Uncontrolled or significant cardiovascular disease including, but not limited to: myocardial infarction or stroke / transient cerebral ischemic attack (TIA) within 6 months prior to consent, Uncontrolled angina within 3 months prior to consent, history of clinically significant arrhythmia (ventricular tachycardia, ventricular fibrillation, or ventricular arrhythmia), QTc prolongation over 480 msec, other clinical History of significant heart disease (ie, cardiomyopathy, congestive heart disease with New York Heart Association [NYHA] functional classification III-IV, pericarditis, significant epicardial fluid).
Requirements related to cardiovascular disease for daily oxygen supplementation.
Confirmed history of encephalitis, meningitis, or uncontrolled seizures prior to informed consent.
Positive blood screen for human immunodeficiency virus (HIV) or known acquired immunodeficiency syndrome (AIDS).
Positive test for hepatitis A antibody (HepA IgM) (note: history of resolved hepatitis A virus infection is not an exclusion criterion), positive for hepatitis B surface antigen (HBsAg) and / or hepatitis B core antigen , A history of any chronic hepatitis evidenced by a positive test for qualitative hepatitis C virus load (by PCR).
Evidence of an active infection requiring systemic antibacterial, antiviral, or antifungal therapy within 7 days prior to initiation of study medication.
Any other significant acute or chronic medical illness.
Subjects who cannot undergo venipuncture and / or cannot tolerate venous access.
Any other legitimate medical, psychiatric, and / or social reasons.
Any of the following procedures or drugs:
Within 2 weeks prior to the time of informed consent: systemic or topical corticosteroids at immunosuppressive doses (7.5 mg / day or more prednisone or equivalent), palliative and gamma knife radiation surgery in the CNS, or medicine Herbal preparation.
Within 4 weeks prior to study drug administration: any study drug or placebo, any anti-cancer therapy (chemotherapy, biologics, therapeutic vaccine, radiation therapy, or hormonal treatment), non-tumor vaccine containing live virus, Allergen desensitization therapy, growth factors such as granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), erythropoietin, major surgery, or bisphosphonate.
Within 10 weeks before administration of test drug: nuclear factor kappa-B ligand receptor activator (RANK-L) inhibitor.

Allergy and adverse drug reactions Anti-PD-1 or anti-PD-L1 antibody therapy, or a history of allergy to other monoclonal antibodies or related compounds or any of their components (eg, against drugs formulated with Polysorbate 80) History of severe hypersensitivity reactions).

Other Exclusion Criteria Prisoners or unwillingly imprisoned subjects.
Subjects who are detained for treatment of a psychiatric or physical (eg, infectious disease) condition.
Failure to comply with restrictions and prohibited activities and actions.

8. Guidelines for Dose Modification Intra-subject dose escalation of BMS-986016 or BMS-936558 is not acceptable in this study. Except for subjects treated as much as possible at dose levels subsequently considered to exceed the MTD, an intra-subject dose reduction of BMS-986016 or BMS-936558 is unacceptable.

  In some cases, the natural history of certain adverse events associated with immunotherapy may be more severe, unlike adverse events caused by other therapeutic classes. Early recognition and management reduces severe toxicity.

In addition, for certain toxicities, management algorithms can be assisted. The toxicity for which the control algorithm was developed includes:
-Lung, gastrointestinal, liver, endocrine, kidney, skin, nerves.

Subjects who experience the following must retain all study drugs:
DLT (by definition, associated with study drug)
• Selected drug-related adverse events and abnormal drug-related laboratory findings:
-Grade 1 or higher pneumonitis-Grade 2 or higher abnormality in AST, ALT, total bilirubin, amylase, or lipase-Grade 2 or higher creatinine-Grade 2 or higher diarrhea or colitis-Grade 2 or higher neurological adverse event / investigation Adverse events, abnormal laboratory findings, or complications that, at the discretion of the investigator, justify delaying administration of the study drug.

  A dosing delay of more than 7 days is considered not to have been administered and is not replaced.

9. Safety Evaluation Adverse events will be evaluated during the study and continuously over 135 days after the last treatment. Adverse events are evaluated according to NCI CTCAE version 4.0. Adverse events are encrypted using the latest version of the Medical Dictionary for Regulatory Activities (MedDRA) and reviewed for potential significance and significance.

10. Other Analysis Depending on the tumor type of interest listed in Table 4 below, various serological tumor markers, gene mutation status, and further analysis are needed. With the exception of serologic tumor markers, if laboratory data from previous studies is available, no assessment is performed.

  Further measurements, including laboratory tests not required for testing, will be performed as clinically indicated. Record the results of all laboratory tests required by this protocol.

11. Efficacy Evaluation Efficacy is evaluated in Parts A and B (dose escalation) and Part C (cohort expansion). The change in tumor measurement and tumor response at the time of each evaluation is determined. Baseline assessment during the screening period requires CT or MRI scans of the chest, abdomen, and pelvis, as well as other anatomical areas, as indicated by the individual subject's tumor type and / or medical history. At subsequent time points, a scan of the chest, abdomen, and pelvis, as well as other anatomical regions, scanned at baseline based on the tumor type and / or history of the individual subject is required. Otherwise, a brain scan is necessary if clinically indicated.

  Analysis of response endpoints indicates that objective and sustained responses reflect clinical experience with other T cell-directed immunotherapy observed in subjects after progression and without intervening alternative anticancer therapies Performed according to response criteria, irRECIST (Wolchok JD, et al., Clin. Can. Res. 2009; 15 (23): 7412-7420). The best overall response (BOR), duration of progression-free survival (PFS), and duration of response (DOR) for each subject are calculated as needed.

  Tumor status is assessed at baseline, during treatment (every 8 weeks) over up to 12 8-week cycles of therapy, and once during follow-up. CT and MRI scans are read again and evaluated locally according to RECIST v1.1. All imaging scans are de-identified and stored as part of the test file of interest in its own Digital Imaging and Communications in Medicine (DICOM) format.

  Efficacy evaluation was performed using irRECIST and RECIST v1.1. Includes ORR (eg, PR + CR), DOR, and PFSR at landmark time (eg, 24 weeks) based on assessment of tumor response using Landmark 2 year overall survival (OS).

12. Pharmacokinetic Evaluation Serum samples for BMS-986016 pharmacokinetic and anti-drug antibody (ADA) evaluation are collected for all subjects. Serum samples for BMS-936558 pharmacokinetics and ADA evaluation will be re-collected for all subjects enrolled in Parts B and C. Serum samples are analyzed for BMS-986016 and BMS-936558 by validated immunoassays. In addition, the selected serum samples are analyzed by an exploratory orthogonal method that measures total BMS-986016 and / or BMS-936558 (eg, liquid chromatography [LC] -mass spectrometry [MS] / MS).

13. Exploratory Biomarker Evaluation The pharmacodynamics of BMS-986016, administered alone or in combination with BMS-936558, was determined for the first three subjects enrolled at each dose level during dose escalation (Parts A and B) and Assess by quantifying biomarkers in peripheral blood and tumor tissue in subjects with melanoma and head and neck cancer during the cohort expansion (Part C) phase of the study. The detailed schedule for pharmacological evaluation is shown in Tables 5-6 below. Details regarding tumor tissue requirements for subjects in Part A, B and C of the study are provided in Table 7 below.

Soluble biomarkers (serum biomarkers)-Part A, B and C
Serum levels of chemokines, cytokines, and tumor-associated soluble proteins before and during treatment are assessed by techniques including, but not limited to, ELISA or multiplex assays. Analytes include markers of inflammation, immune activation, host tumor growth factors, and tumor-derived proteins.

Anti-tumor antibodies (serum biomarkers)-Part A, B and C
Treatment with BMS-986016 and BMS-936558 may result in the production of new antibodies against tumor-associated antigens, or an increase in the antibodies present. Evaluation of antibodies against a panel of more than 8000 proteins is performed using sera before and during treatment in multiplex assays and ELISAs. These data are used to explore whether anti-tumor antibodies are associated with clinical response and safety parameters, and to provide information on the pharmacodynamics of drug administration.

Immunophenotyping (flow cytometry / PBMC)-Part A, B and C
Peripheral blood mononuclear cells (PBMC) are used to characterize and quantify the activation and regulatory status of myeloid and lymphoid cells by multicolor flow cytometry. The subset of cells that are characterized by immunophenotyping include na 、 ve, activated and depleted effector and memory T cell populations, regulatory T cells, and bone marrow-derived suppressor cells.

Ex Vivo Functional Assay (Cellular Assay)-Part A, B and C
PBMCs are isolated and cryopreserved to assess whether BMS-986016 and BMS-936558 restore T cell activation and function. The functional status of effector T cells such as, but not limited to, IFN-γ and granzyme B is assessed by flow cytometric staining.

Peripheral blood gene expression (whole blood mRNA) and oncogene expression-Part A, B and C
Expression levels of genes associated with response to BMS-986016 ± BMS-936558 are quantified by microarray and / or quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis in whole blood and tumor samples. Analyzes include, but are not necessarily limited to, genes encoding effector functions (perforin, granzyme B, and IFN-γ) stimulated by BMS-986016 and T cell costimulatory receptors (PD-1, PD -L1 and CTLA-4).

Circulating tumor DNA analysis (serum (plasma) biomarkers)-Part C
The presence of cell-free DNA in the circulating blood is a well-documented phenomenon. Fragments of DNA flow from dividing cells into the bloodstream during cell growth or cell death. In patients with cancer, this DNA fraction is of tumor origin and is called circulating tumor DNA (ctDNA). Despite being small, average 180-200 bp fragments of DNA and specific genomic regions can be amplified using PCR. In addition, several studies have detected mutations in ctDNA that exactly match those from the parent tumor. Using tissue and plasma from patients with known driver mutations in melanoma or head and neck cancer, the frequency of mutations in circulation is counted using the BEAMing technique.

Single nucleotide polymorphism analysis (SNP)-Part A, B and C
The genes selected for single nucleotide polymorphisms (SNPs) are evaluated to identify potential polymorphisms associated with the safety and efficacy of BMS-986016. Genes of interest include, but are not limited to, PD-1, PD-L1, MHC class II, LAG-3, and CTLA-4.

Tumor biopsy analysis-Part A and B
Tumor tissue is collected from all subjects during the dose escalation portion of the protocol. The number and composition of immunoinfiltrates are evaluated using immunohistochemical analysis to define a subset of immune cells present in FFPE tumor tissue before and potentially after exposure to BMS-986016 and BMS-936558 . These IHC analyzes include, but are not necessarily limited to, the following markers: CD4, CD8, LAG-3, MHCII, PD-1, PD-L1, and PD-L2. A correlation between gene expression and IHC expression is made between assays performed when deemed beneficial.

Tumor-based biomarker measurement-Part C
Paired pre-treatment and intra-treatment tumor biopsies are mandatory for all subjects with melanoma or head and neck cancer enrolled in Part C (Cohort Expansion). A well-paired pre-procedure and intra-procedure biopsy can replace subjects whose biopsies are not obtained.

  The subject has at least one lesion large enough to repeatedly undergo biopsy (pre-procedure and intra-procedure biopsy) with a core needle (minimum size 18 gauge), or a needle or resection biopsy Having at least two different lesions that are eligible. Expected core needle length should be greater than 5 mm. Punch biopsy is acceptable in skin lesions. Fine needle aspiration biopsy is not allowed. At least two core biopsies will be obtained at each time point, but if deemed clinically safe by the Investigator, further core collection is strongly encouraged. Assessment of biopsy quality by a pathologist is strongly encouraged at the time of the procedure. For all biopsies taken, a detailed pathology report must be submitted with the specimen.

Tumor biopsy specimens are obtained from subjects who agree before and during treatment with BMS-986016 and BMS-936558 to characterize the immune cell population and expression of selected tumor markers. Biopsy samples are used for the following evaluations:
-Characterization of TIL and tumor antigen. The number and composition of immune infiltrates is evaluated using immunohistochemical analysis to define the subset of immune cells present in FFPE tumor tissue before and after exposure to BMS-986016 and nivolumab. These IHC analyzes include, but are not necessarily limited to, the following markers: CD4, CD8, LAG-3, MHCII, PD-1, PD-L1, and PD-L2. A correlation between gene expression and IHC expression is made between assays performed when deemed beneficial.
・ Laser capture microdissection. Isolation of tumor and / or TIL on FFPE sections is performed by laser capture microdissection (LCM) for efficient profiling of molecular events within the tumor microenvironment.
-Characterization of the T cell repertoire. DNA sequencing is performed on FFPE tumor tissue before and after treatment to assess the composition of the T cell repertoire. Low T cell receptor diversity may be a poor prognostic factor for overall survival in metastatic breast cancer patients. Given that it is currently postulated that the major mechanism of BMS-936558 and BMS-986016 is the functional recovery of T-cell antitumor immunity, T-cell receptor diversity as a predictor of response to immunotherapy is I don't know. Therefore, characterization of the diversity of the T cell compartment at baseline and in the periphery during treatment and within the tumor is performed by T cell receptor next generation DNA sequencing. Also, T cell repertoire analysis is performed from DNA isolated from peripheral blood to compare the status of the tumor and peripheral T cell repertoire before and after treatment.
-Gene expression profiling. Tumor biopsies taken in RNAlater or similar reagents are tested for mRNA gene expression by Affymetrix gene array technology and / or quantitative real-time polymerase chain reaction (qPCR) to detect expression of selected immune-related genes I do.
-Expression of in situ cytokines and negative regulators. Tumor biopsies are evaluated quantitatively for RNA, including CD3, IFN-γ, LAG-3, and PD-1.

  Subjects with insufficient tissue quality or quality from screening biopsies will continue testing. If the biopsy during the procedure is not successful, the subject will continue the study. Replacing such subjects will yield 48 subjects with sufficient tumor biopsy pairs. A biopsy during the procedure may not be possible if the subject has a response to the procedure. In this case, the subject will also continue the test.

  Tumor tissue from these biopsies can be split into equal parts into FFPE and frozen samples and used for histological confirmation of melanoma and the assays listed above.

  Biopsies are performed using local anesthesia or conscious sedation. Follow institutional guidelines for safe biopsy performance. An excisional biopsy is performed to obtain a tumor biopsy. Invasive procedures requiring general anesthesia are not performed to obtain biopsy specimens. However, when performing surgical procedures for clinical indications, with the consent of the subject, excess tumor tissue is used for research purposes.

14． Immunogenicity assessment Serum samples taken at each time point are analyzed by a validated immunogenicity assay. Selected serum samples are analyzed by an exploratory orthogonal method that measures anti-BMS-986016 or anti-BMS-936558. Potential results generated from any orthogonal method are intended to be useful for technology exploration purposes and will not be re-reported.

  In addition, ad hoc serum samples designated for pharmacokinetic or biomarker evaluation are used for immunogenicity analysis as needed (eg, for complete immunogenicity assessment or associated with suspected immunogenicity). Not enough capacity to track adverse events).

15. Adverse Events Adverse events (AEs) are defined as any new adverse medical events or exacerbations of the pre-existing medical condition in clinical subjects who are administered the investigational (medicine) drug and do not necessarily have a causal relationship to this treatment. Defined. Thus, an AE is any undesired and unintended sign (such as an abnormal laboratory finding), symptom, or disease that is temporally associated with the use of an investigational drug, whether or not it is considered to be associated with the investigational drug. .

The causal relationship to the study drug is determined by the physician and is used to assess any adverse events (AEs). The causal relationship may be one of the following:
Association: There is a rational causal relationship between test drug administration and AE.
Unrelated: There is no rational causal relationship between study drug administration and AE.

  The term "rational causality" means that there is evidence to suggest a causality.

Serious Adverse Events Serious adverse events (SAEs)
• causes death • is life-threatening (defined as an event in which the subject was at risk of death at the time of the event; it is assumed that if it were more severe, it caused death) Does not mean)
• requires hospitalization or causes prolonged hospitalization that has already begun; • results in persistent or significant disability / inability; • is a birth defect / birth defect. Does not threaten or cause death or hospitalization, but based on sound medical and scientific judgment, puts the subject at risk or other serious consequences listed in the above definitions Is any deleterious medical event that is defined as a medical event that may require intervention (eg, medical, surgical) to prevent one of the following. Examples of such events include, but are not limited to, intensive care in an emergency room or at home for allergic bronchospasm; a blood disease or convulsion that does not result in hospitalization. Potential drug-induced liver injury (DILI) is also considered an important medical event.

  A suspected transmission of the infectious agent (eg, pathogenic or non-pathogenic) by the test drug is SAE. According to regulatory definitions, pregnancy, overdose, cancer, and potential drug-induced liver injury (DILI) are not always serious, but these events must be treated as SAEs. Any component of the study endpoint that is considered relevant to the study therapy (eg, death is the endpoint and if mortality occurs due to anaphylaxis, anaphylaxis must be reported) is reported as SAE.

The following hospitalizations are not considered SAEs:
-Visits to an emergency room or other medical department for less than 24 hours that do not result in hospitalization (unless considered a significant medical or life-threatening event).
-Elective surgery planned before signing consent.
-Admission according to protocol for planned medical / surgical procedures.
-Routine health assessments requiring hospitalization for baseline / trends in health status (eg routine colonoscopy).
-Medical / surgical procedures other than to improve illness and planned before enrollment in the trial. Proper documentation is required in these cases.
-Due to alternative living conditions that do not affect health and do not require medical / surgical intervention (eg lack of housing, financial inadequacy, caregiver cessation, family environment, administrative reasons). Hospitalization.

  Following the written consent of the subjects participating in the study, all SAEs, whether related to the study drug or not, are collected, including those considered to be related to protocol-specific procedures. All SAEs that occur within the screening period and within 135 days of discontinuation of dosing will be collected. If applicable, collect the SAE associated with it for any subsequent protocol-specific procedures (eg, follow-up skin biopsy). All SAEs are tracked until resolution or stabilization.

Non-serious adverse events Non-serious adverse events are AEs that are not classified as serious. Collection of non-serious AE information begins at the start of study drug and lasts 135 days after discontinuation of dosing. Non-serious AEs are followed until resolution or stabilization, or if they become severe, they are reported as SAEs. Follow-up is required for non-severe AEs that cause discontinuation or discontinuation of study drug, as well as those present at the end of study treatment as appropriate. All identified non-serious AEs are recorded and described on the CRF (paper or electronic) non-serious AE page.

  Require additional CRFs to be completed for AEs and / or laboratory abnormalities reported / identified during the course of the study.

16. Statistical considerations Sample size determination Dose escalation (Parts A and B): The sample size at each dose depends on the toxicity observed and cannot be determined accurately. Part A and Part B have 3-9 subjects in each cohort.

  Cohort expansion (Part C): Cohort expansion allows for better estimates of toxicity rates and provides better accuracy close to preliminary estimates of efficacy. If less than 5 of 16 subjects (ie, about 30% of the cohort) experience toxicity, there is at least 90% confidence that the true rate of toxicity is less than 50.4% ( (Based on Chopper-Pearson direct binomial one-sided 90% confidence interval). The sample size of the 16 subjects per cohort also indicates an objective response such that the maximum distance between the estimate and any of the limits of the direct 2-sided 95% Chopper-Pearson confidence interval is 27.4%. In the cohort (ie, CR + PR).

Population / all enrollment analysis set for analysis: This analysis set contains all subjects (including screening failures) that have signed informed consent for the study.
• All treatments analysis set: This analysis set includes all subjects receiving any drug.
-Response evaluable subject: This analysis set receives any test drug, baseline tumor assessment for measurable disease, and: (1) at least one evaluable tumor assessment during treatment; (2) Includes all subjects with one of clinical progression or (3) death before tumor assessment during initial treatment.
BMS-986016 pharmacokinetic analysis set: This analysis set includes all subjects who have received BMS-9886016 and have at least one valid PK parameter included in the statistical analysis of the BMS-9886016 PK data.
-BMS-986016 immunogenicity analysis set: This analysis set includes all subjects who have received BMS-9886016 and have at least one available BMS-986016 immunogenic sample.
BMS-98558 immunogenicity analysis set: This analysis set includes all subjects who have received BMS-98558 and have at least one available BMS-98558 immunogenic sample.
• Pharmacodynamic analysis set: This analysis set includes all treated subjects for whom pharmacodynamic measurements are available at baseline and at least one other time point.

Endpoints The primary endpoints of this phase 1 study are AEs, serious adverse events (SAEs), deaths, and abnormal laboratory findings (e.g., according to (Grade 3 or higher). All subjects receiving at least one dose of BMS-986016 or BMS-936558 will be analyzed for safety.

PK of BMS-986016 administered alone or in combination with BMS-936558 is evaluated as a secondary objective using the following endpoints derived from serum concentration versus time data in Cycle 1 and Cycle 3: :
Cmax Maximum blood concentration Tmax Time to reach maximum blood concentration Ctrough Trough blood concentration Ctau Concentration at end of dosing interval (eg, concentration at 336 hours)
Css, avg Average concentration over dosing interval ([AUC (TAU) / Tau])
AUC (TAU) Area under the concentration time curve at one dosing interval CLT Whole body clearance Vss Distribution volume at steady state T-HALFeff AUC Effective elimination half-life T-HALFeff Cmax explaining observed AUC accumulation Degree Cmax accumulation observed Effective elimination half-life, AI_AUC, describing the degree; Accumulation index; ratio of AUC (TAU) at steady state to AUC (TAU) after first administration AI_Cmax Cmax accumulation index; Cmax at steady state, Cmax after first administration Specific AI_Ctau Ctau accumulation index; ratio DF variability or variability index between Ctau in the steady state and Ctau after the first administration ([Cmax-Ctau] / Css, avg).

  Individual subject PK parameter values are derived by a non-compartmental method with a validated PK analysis program. The actual time is used for analysis.

Indications Efficacy is evaluated as a secondary objective using the endpoints described below for irRECIST and RECIST v1.1. For patient management, clinical decision making is based on RECIST. Statistical analysis and reporting are based on both criteria.
Best overall effect (BOR) is based on RECIST v1.1 or irRECIST criteria, taking into account the requirements for confirmation and the last protocol-specific tumor assessment from the start of study treatment (eg 30 day follow-up visit) Means the best response recorded. The CR or PR determination included in the BOR assessment is confirmed by two consecutive (confirmative) assessments that meet the criteria for response and is performed at least four weeks after the criteria for response are first met.
Objective Response Rate (ORR) is defined as the total number of subjects whose BOR is either CR or PR divided by the total number of subjects in the subject population.
The duration of response (DOR), calculated only for subjects with a BOR of CR or PR, whichever occurs first, is objectively documented based on the date of the first response and criteria (RECISTv1.1 or irRECIST) It is defined as the number of days between the later date of the disease progression or death taken. For subjects who are still alive, in progress, or have not received any subsequent therapy, the duration of response is terminated at the date of the last protocol-specific tumor assessment. Subjects receiving subsequent therapy will be discontinued at the start of subsequent therapy.
Progression free survival (PFS) is defined as the probability of a subject being still progression and surviving. This probability is calculated based on the number of days between the first dose of study drug and progressive disease (defined by RECIST or irRECIST) or death. For those who are still alive and in progress, PFS will be censored at the date of the last protocol-specific tumor assessment.

  These endpoints were for tumor measurements performed every 8 weeks (wefek) during the treatment period (up to 12 8-week cycles) and once during the clinical follow-up period (30 days) for a total of about 1.9 years. It is determined based on.

Immunogenicity At the sample level, individual samples are characterized as ADA positive or ADA negative. A subject is considered to have a positive sample at baseline if the last sample before the start of treatment is ADA positive. For example, a post-baseline sample from a subject that is ADA negative at baseline is considered ADA positive if ADA is detected. A post-baseline sample from a subject that is ADA positive at baseline is considered ADA positive if there is an associated titer increase (the magnitude of the titer increase considered relevant depends on the drug and assay). May vary and is indicated in the statistical analysis plan). At the target level, the relevant ADA endpoints are:
-Proportion of subjects showing ADA positive samples at baseline-Proportion of ADA positive controls (during treatment and overall)
The percentage of subjects that are persistently positive (eg, two or more consecutive ADA-positive samples during which time has elapsed sufficiently) the percentage of subjects in which neutralizing antibodies are detected in one or more samples May be included.

ECG read in center (parts A and B)
In Part A and Part B, QTc is assessed by a central reader at follow-up visit 1 and on day 1 of cycle 1 and cycle 3 (pre-dose and 4 hours post-dose). These assessments are used to address the secondary purpose of assessing the effects of BMS-986016, administered alone and in combination with BMS-936558, on QTc. An ECG evaluated locally by the investigator is also taken at the beginning of each cycle.

Biomarker endpoints Biomarker endpoints from peripheral blood are generally measured at multiple time points and evaluated as both predictive and pharmacodynamic markers in the context of exploratory biomarker goals. These may include measures such as the level of the following at each planned point in time and the change from baseline at that level:
• Serum soluble factor • Specific lymphocyte subset / expression level ratio of T cell costimulatory markers assessed using flow cytometry • Effector functions stimulated by BMS-986016 (perforin, granzyme B, and IFN-γ ) And expression of genes encoding T cell costimulatory receptors (PD-1, PD-L1, and CTLA-4). For expression of a single nucleotide polymorphism associated with the PD-1 gene Percentage (by SNP)
A measure of the amount and diversity of antibodies observed against tumor-associated antigens (Part C only).

Because biomarker endpoints from tumor biopsies are measured only at baseline for many subjects, they are primarily searched to identify baseline markers that predict efficacy. The pharmacodynamic relationships are explored for a subset of subjects who have both pre-treatment and intra-treatment biopsies. Evaluation items are
• Percentage of CD8 + T cells that are positive for IFN-γ and granzyme B expression, and functional status of lymphocytes (ex vivo) measured as geometric mean intensity (log scale) of CD8 + cells that are positive for IFN-γ and granzyme B expression Functional assay)
-Genes encoding effector functions stimulated by BMS-986016 (perforin, granzyme B, and IFN-γ) and genes encoding T cell costimulatory receptors (PD-1, PD-L1, and CTLA-4) Expression of LAG-3, MHC class II, PD-1, PD-L1, and PD-L2 expression presence and intensity (measured using individual scales such as 0, 1, 2, 3, 4, etc.) ) May include measures such as pre-treatment levels of IHC assessment and changes in levels observed during treatment.

  Appropriate functional transformations of these exploratory measures are applied as needed.

Pharmacokinetics Planned troughs and BMS-936558 concentration-time data at the end of the infusion are evaluated as exploratory endpoints. Measurements are collected during treatment (up to 12 cycles) and up to 135 days during follow-up after treatment.

  The PK parameters for BMS-986016 are calculated using non-compartmental analysis. Summary statistics are tabulated for PK parameters of BMS-986016 by dose and test date / cycle. To describe the relationship between these parameters and the dose of BMS-986016, a scatter plot of Cmax and AUC (TAU) versus dose is provided for each day / cycle measured. The dose proportionality of BMS-986016, administered alone or co-administered with BMS-936558, is also evaluated based on a power model. The trough concentration of BMS-986016 is plotted again against test date and cycle. BMS-936558 infusion end and trough concentrations are tabulated using summary statistics.

  Array overview

Claims (23)

A method of treating a solid tumor in a human patient, comprising:
(A) an anti-LAG- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 5; 3 antibodies,
(B) an anti-PD- comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence of SEQ ID NO: 19, and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence of SEQ ID NO: 21; Administering an effective amount of each of the one antibody,
Including at least one dosing cycle, the cycle is eight weeks, and for at least one of each cycle, four doses of anti-LAG-3 antibody are administered at a dose of 3, 20, 80, or 240 mg, and four doses of anti-PD- The method, wherein one antibody is administered at a dose of 80 or 240 mg. An anti-LAG-3 antibody and an anti-PD-1 antibody
(A) 3 mg of anti-LAG-3 antibody and 80 mg of anti-PD-1 antibody;
(B) 3 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
(C) 20 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody;
2. The method according to claim 1, wherein the agent is administered at a dose of (d) 80 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody; or (e) 240 mg of anti-LAG-3 antibody and 240 mg of anti-PD-1 antibody. the method of.   2. The method of claim 1, wherein the anti-PD-1 and anti-LAG-3 antibodies are formulated for intravenous administration.   2. The method of claim 1, wherein the anti-PD-1 and anti-LAG-3 antibodies are co-formulated.   The method according to claim 1, wherein the anti-PD-1 antibody and the anti-LAG-3 antibody are separately formulated.   2. The method of claim 1, wherein the treatment comprises up to 12 cycles.   2. The method of claim 1, wherein the anti-PD-1 antibody is administered on days 1, 15, 29 and 43 of each cycle.   2. The method of claim 1, wherein the anti-LAG-3 antibody is administered on days 1, 15, 29 and 43 of each cycle.   2. The method of claim 1, wherein the anti-PD-1 antibody is administered prior to the administration of the anti-LAG-3 antibody.   10. The method of claim 9, wherein the anti-LAG-3 antibody is administered within about 30 minutes before administration of the anti-PD-1 antibody.   2. The method of claim 1, wherein the anti-PD-1 antibody is administered after administration of the anti-LAG-3 antibody.   2. The method of claim 1, wherein the anti-PD-1 antibody is administered simultaneously with the anti-LAG-3 antibody.   2. The method of claim 1, wherein the treatment results in at least one therapeutic effect selected from a reduction in tumor size, a reduction in the number of metastatic lesions over time, a complete response, a partial response, and a stable disease.   2. The method of claim 1, wherein the solid tumor is selected from melanoma, non-small cell lung cancer (NSCLC), human papillomavirus (HPV) -related tumor, and gastric adenocarcinoma. An anti-LAG-3 antibody
(A) a heavy chain variable region CDR1 comprising the sequence of SEQ ID NO: 7;
(B) a heavy chain variable region CDR2 comprising the sequence of SEQ ID NO: 8;
(C) a heavy chain variable region CDR3 comprising the sequence of SEQ ID NO: 9;
(D) a light chain variable region CDR1 comprising the sequence of SEQ ID NO: 10;
(E) a light chain variable region CDR2 comprising the sequence of SEQ ID NO: 11; and (f) a light chain variable region CDR3 comprising the sequence of SEQ ID NO: 12.
The method of claim 1, comprising:   2. The method of claim 1, wherein the anti-LAG-3 antibody comprises a heavy and light chain variable region comprising the sequences set forth in SEQ ID NO: 3 and SEQ ID NO: 5, respectively.   2. The method of claim 1, wherein the anti-LAG-3 antibody comprises a heavy and light chain comprising the sequence set forth in SEQ ID NO: 1 and SEQ ID NO: 2, respectively. An anti-PD-1 antibody
(A) a heavy chain variable region CDR1 comprising the sequence of SEQ ID NO: 23;
(B) a heavy chain variable region CDR2 comprising the sequence of SEQ ID NO: 24;
(C) a heavy chain variable region CDR3 comprising the sequence of SEQ ID NO: 25;
(D) a light chain variable region CDR1 comprising the sequence of SEQ ID NO: 26;
(E) a light chain variable region CDR2 comprising the sequence of SEQ ID NO: 27; and (f) a light chain variable region CDR3 comprising the sequence of SEQ ID NO: 28
The method of claim 1, comprising:   2. The method of claim 1, wherein the anti-PD-1 antibody comprises a heavy and light chain variable region comprising the sequences set forth in SEQ ID NO: 19 and SEQ ID NO: 21, respectively.   The method of claim 1, wherein the anti-PD-1 antibody comprises a heavy and light chain comprising the sequences set forth in SEQ ID NO: 17 and SEQ ID NO: 18, respectively. A kit for treating a solid tumor in a human patient, comprising:
(A) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 5 Anti-LAG-3 antibody;
(B) one dose comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 21 A kit comprising: an anti-PD-1 antibody; and (c) instructions for using the anti-LAG-3 antibody and the anti-PD-1 antibody in the method of claim 1.   In at least one cycle, an antibody comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 21. CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3 and CDR1, CDR2 of the light chain variable region having the sequence set forth in SEQ ID NO: 5 for co-administration with a PD-1 antibody And a CDR3 domain comprising four doses of anti-LAG-3 antibody at each dose of 3, 20, 80 or 240 mg and four doses of anti-PD-1 antibody for each cycle. Alternatively, an anti-LAG-3 antibody administered at a dose of 240 mg.   In at least one cycle, an antibody comprising the CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 19 and the CDR1, CDR2 and CDR3 domains of the light chain variable region having the sequence set forth in SEQ ID NO: 21. CDR1, CDR2 and CDR3 domains of the heavy chain variable region having the sequence set forth in SEQ ID NO: 3 and CDR1, CDR2 of the light chain variable region having the sequence set forth in SEQ ID NO: 5 for co-administration with a PD-1 antibody Anti-LAG-3 antibody comprising an anti-PD-1 antibody and a CDR3 domain, wherein for each cycle four doses of the anti-LAG-3 antibody are administered at a dose of 3, 20, 80 or 240 mg / kg, and four doses of the anti-PD-1 antibody Is administered at a dose of 80 or 240 mg / kg.

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